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Review ArticleLipid Peroxidation Production Metabolism and SignalingMechanisms of Malondialdehyde and 4-Hydroxy-2-Nonenal

Antonio Ayala Mario F Muntildeoz and Sandro Arguumlelles

Department of Biochemistry and Molecular Biology Faculty of Pharmacy University of SevilleProf Garcıa Gonzales sn 41012 Seville Spain

Correspondence should be addressed to Sandro Arguelles arcasanalumuses

Received 14 February 2014 Accepted 24 March 2014 Published 8 May 2014

Academic Editor Kota V Ramana

Copyright copy 2014 Antonio Ayala et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Lipid peroxidation can be described generally as a process under which oxidants such as free radicals attack lipids containingcarbon-carbon double bond(s) especially polyunsaturated fatty acids (PUFAs) Over the last four decades an extensive body ofliterature regarding lipid peroxidation has shown its important role in cell biology and human health Since the early 1970s the totalpublished research articles on the topic of lipid peroxidation was 98 (1970ndash1974) and has been increasing at almost 135-fold by up to13165 in last 4 years (2010ndash2013) New discoveries about the involvement in cellular physiology and pathology as well as the controlof lipid peroxidation continue to emerge every day Given the enormity of this field this review focuses on biochemical concepts oflipid peroxidation productionmetabolism and signalingmechanisms of twomain omega-6 fatty acids lipid peroxidation productsmalondialdehyde (MDA) and in particular 4-hydroxy-2-nonenal (4-HNE) summarizing not only its physiological and protectivefunction as signaling molecule stimulating gene expression and cell survival but also its cytotoxic role inhibiting gene expressionand promoting cell death Finally overviews of in vivomammalian model systems used to study the lipid peroxidation process andcommon pathological processes linked to MDA and 4-HNE are shown

This review paper is dedicated toDr Alberto Machado

1 Lipids Overview of Biological Functions

Lipids Are Classically Divided into Two Groups Apolar andPolar Triglycerides (apolar) stored in various cells butespecially in adipose (fat) tissue are usually the main form ofenergy storage in mammals [1 2] Polar lipids are structuralcomponents of cell membranes where they participate in theformation of the permeability barrier of cells and subcellularorganelles in the form of a lipid bilayer The major lipid typedefining this bilayer in almost all membranes is glycerol-based phospholipid [3] The importance of the membranelipid physical (phase) state is evidenced by the fact that lipidsmay control the physiological state of a membrane organelleby modifying its biophysical aspects such as the polarity

and permeability Lipids also have a key role in biology assignaling molecules

Lipids as SignalingMoleculesThemain enzymes that generatelipid signaling mediators are lipoxygenase which medi-ate hydroperoxyeicosatetraenoic acids (HPETEs) lipoxinsleukotrienes or hepoxilins biosynthesis after oxidation ofarachidonic acid (AA) [4 5] cyclooxygenase that producesprostaglandins [4] and cytochrome P-450 (CYP) whichgenerates epoxyeicosatrienoic acids leukotoxins thrombox-ane or prostacyclin [4] Lipid signaling may occur viaactivation of a variety of receptors including G protein-coupled and nuclear receptors Members of several differentlipid categories have been identified as potent intracellularsignal transduction molecules Examples of signaling lipids

Hindawi Publishing CorporationOxidative Medicine and Cellular LongevityVolume 2014 Article ID 360438 31 pageshttpdxdoiorg1011552014360438

2 Oxidative Medicine and Cellular Longevity

include (i) two derived from the phosphatidylinositol phos-phates diacylglycerol (DAG) and inositol phosphates (IPs)DAG is a physiological activator of protein kinase C [6 7]and transcription factor nuclear factor-kB (NF-120581B) whichpromotes cell survival and proliferation Diacylglycerol alsointeracts indirectly with other signalling molecules such assmall G proteins [8] IPs are a highly charged family oflipid-derived metabolites involved in signal transductionthat results in activation of Akt mTOR [9] and calcium-homeostasis [10 11] (ii) sphingosine-1-phosphate a sph-ingolipid derived from ceramide that is a potent messen-ger molecule involved in regulating calcium mobilizationmigration adhesion and proliferation [12ndash14] (iii) theprostaglandins which are one type of fatty-acid derivedeicosanoid involved in inflammation [15 16] and immunity[17] (iv) phosphatidylserine a phospholipid that plays animportant role in a number of signaling pathways includeskinases small GTPases and fusogenic proteins [18] (v) thesteroid hormones such as estrogen testosterone and cortisolwhich modulate a host of functions such as reproductionmetabolism stress response inflammation blood pressureand salt and water balance [19]

2 Lipids Damage by Reactive Oxygen Species

One of the consequences of uncontrolled oxidative stress(imbalance between the prooxidant and antioxidant levelsin favor of prooxidants) is cells tissues and organs injurycaused by oxidative damage It has long been recognized thathigh levels of free radicals or reactive oxygen species (ROS)can inflict direct damage to lipids The primary sources ofendogenous ROS production are the mitochondria plasmamembrane endoplasmic reticulum and peroxisomes [20]through a variety of mechanisms including enzymatic reac-tions andor autooxidation of several compounds suchas catecholamines and hydroquinone Different exogenousstimuli such as the ionizing radiation ultraviolet raystobacco smoke pathogen infections environmental toxinsand exposure to herbicideinsecticides are sources of in vivoROS production

The two most prevalent ROS that can affect profoundlythe lipids are mainly hydroxyl radical (HO∙) and hydroper-oxyl (HO∙

2

) The hydroxyl radical (HO∙) is a small highlymobile water-soluble and chemically most reactive speciesof activated oxygen This short-lived molecule can be pro-duced from O

2

in cell metabolism and under a variety ofstress conditions A cell produces around 50 hydroxyl radicalsevery second In a full day each cell would generate 4million hydroxyl radicals which can be neutralized or attackbiomolecules [21] Hydroxyl radicals cause oxidative damageto cells because they unspecifically attack biomolecules [22]located less than a few nanometres from its site of generationand are involved in cellular disorders such as neurodegenera-tion [23 24] cardiovascular disease [25] and cancer [26 27]It is generally assumed that HO∙ in biological systems isformed through redox cycling by Fenton reaction where freeiron (Fe2+) reacts with hydrogen peroxide (H

2

O2

) and theHaber-Weiss reaction that results in the production of Fe2+

Haber-Weiss reaction

Fenton reaction

O2

∙OH + H2OH2O2

H+

H+ M(n+1) Mn

O2∙minus

HO∙2

Figure 1 Fenton and Haber-Weiss reaction Reduced form oftransition-metals (M119899) reacts trough the Fenton reaction withhydrogen peroxide (H

2

O2

) leading to the generation of ∙OHSuperoxide radical (O

2

∙minus) can also react with oxidized form oftransitionmetals (M(119899+1)) in the Haber-Weiss reaction leading to theproduction of M119899 which then again affects redox cycling

when superoxide reacts with ferric iron (Fe3+) In additionto the iron redox cycling described above also a number ofother transition-metal including Cu Ni Co and V can beresponsible for HO∙ formation in living cells (Figure 1)

The hydroperoxyl radical (HO∙2

) plays an importantrole in the chemistry of lipid peroxidation This protonatedform of superoxide yields H

2

O2

which can react with redoxactive metals including iron or copper to further generateHO∙ through Fenton or Haber-Weiss reactions The HO∙

2

is a much stronger oxidant than superoxide anion-radicaland could initiate the chain oxidation of polyunsaturatedphospholipids thus leading to impairment of membranefunction [28ndash30]

21 Lipid Peroxidation Process Lipid peroxidation can bedescribed generally as a process under which oxidants suchas free radicals or nonradical species attack lipids containingcarbon-carbon double bond(s) especially polyunsaturatedfatty acids (PUFAs) that involve hydrogen abstraction froma carbon with oxygen insertion resulting in lipid per-oxyl radicals and hydroperoxides as described previously[31] Glycolipids phospholipids (PLs) and cholesterol (Ch)are also well-known targets of damaging and potentiallylethal peroxidative modification Lipids also can be oxi-dized by enzymes like lipoxygenases cyclooxygenases andcytochrome P450 (see above lipid as signaling molecules)In response to membrane lipid peroxidation and accord-ing to specific cellular metabolic circumstances and repaircapacities the cells may promote cell survival or inducecell death Under physiological or low lipid peroxidationrates (subtoxic conditions) the cells stimulate their mainte-nance and survival through constitutive antioxidants defensesystems or signaling pathways activation that upregulateantioxidants proteins resulting in an adaptive stress responseBy contrast under medium or high lipid peroxidation rates(toxic conditions) the extent of oxidative damage overwhelmsrepair capacity and the cells induce apoptosis or necrosisprogrammed cell death both processes eventually lead tomolecular cell damage which may facilitate development ofvarious pathological states and accelerated agingThe impactof lipids oxidation in cell membrane and how these oxidativedamages are involved in both physiological processes andmajor pathological conditions have been analysed in severalreviews [32ndash35]

Oxidative Medicine and Cellular Longevity 3

The overall process of lipid peroxidation consists of threesteps initiation propagation and termination [31 36 37]In the lipid peroxidation initiation step prooxidants likehydroxyl radical abstract the allylic hydrogen forming thecarbon-centered lipid radical (L∙) In the propagation phaselipid radical (L∙) rapidly reacts with oxygen to form a lipidperoxy radical (LOO∙) which abstracts a hydrogen fromanother lipid molecule generating a new L∙ (that continuesthe chain reaction) and lipid hydroperoxide (LOOH) In thetermination reaction antioxidants like vitamin E donate ahydrogen atom to the LOO∙ species and form a correspond-ing vitamin E radical that reacts with another LOO∙ formingnonradical products (Figure 2) Once lipid peroxidation isinitiated a propagation of chain reactions will take place untiltermination products are produced Review with extensiveinformation regarding the chemistry associated with each ofthese steps is available [31]

22 Lipid Peroxidation Products Lipid peroxidation or reac-tion of oxygen with unsaturated lipids produces a widevariety of oxidation products The main primary productsof lipid peroxidation are lipid hydroperoxides (LOOH)Among the many different aldehydes which can be formed assecondary products during lipid peroxidation malondialde-hyde (MDA) propanal hexanal and 4-hydroxynonenal (4-HNE) have been extensively studied by Esterbauer and hiscolleagues in the 80s [38ndash49] MDA appears to be the mostmutagenic product of lipid peroxidation whereas 4-HNE isthe most toxic [50]

MDAhas beenwidely used formany years as a convenientbiomarker for lipid peroxidation of omega-3 and omega-6fatty acids because of its facile reaction with thiobarbituricacid (TBA) [48 51] The TBA test is predicated upon thereactivity of TBA toward MDA to yield an intensely coloredchromogen fluorescent red adduct this test was first used byfood chemists to evaluate autoxidative degradation of fats andoils [52] However the thiobarbituric acid reacting substancestest (TBARS) is notoriously nonspecific which has led tosubstantial controversy over its use for quantification ofMDAfrom in vivo samples Several technologies for the determi-nation of free and total MDA such gas chromatography-mass spectrometry (GC-MSMS) liquid chromatography-mass spectrometry (LC-MSMS) and several derivatization-based strategies have been developed during the last decade[53] Because MDA is one of the most popular and reliablemarkers that determine oxidative stress in clinical situations[53] and due toMDArsquos high reactivity and toxicity underlyingthe fact that this molecule is very relevant to biomedicalresearch community

4-HNE was first discovered in 60s [54] Later in 80s4-HNE was reported as a cytotoxic product originatingfrom the peroxidation of liver microsomal lipids [40] 4-Hydroxyalkenals produced in the course of biomembranelipids peroxidation elicited either by free radicals or bychemicals might exert a genotoxic effect in humans [55]The4-hydroxyalkenals are the most significant products becausethey are produced in relatively large amounts and they arevery reactive aldehydes that act as ldquosecond messengers of

free radicalsrdquo In particular 4-HNE which has been subjectedto intense scientific scrutiny in 90s [49] is consideredas ldquoone of the major toxic products generated from lipidperoxidesrdquo [49] 4-HNE high toxicity can be explained by itsrapid reactions with thiols and amino groups [56] Reactivealdehydes especially 4-HNE act both as signaling molecules(see below 4-HNE as signaling molecule) and as cytotoxicproducts of lipid peroxidation causing long-lasting biologicalconsequences in particular by covalent modification ofmacromolecules (see below 4-HNE biomolecular adducts)4-HNE is considered as ldquosecond toxic messengers of freeradicalsrdquo and also as ldquoone of the most physiologically activelipid peroxidesrdquo ldquoone of major generators of oxidative stressrdquoldquoa chemotactic aldehydic end-product of lipid peroxidationrdquoand a ldquomajor lipid peroxidation productrdquo [57] Thus itis not a surprise that 4-HNE is nowadays considered asmajor bioactive marker of lipid peroxidation and a signalingmolecule involved in regulation of several transcriptionfactors sensible to stress such as nuclear factor erythroid2-related factor 2 (Nrf2) activating protein-1 (AP-1) NF-120581B and peroxisome-proliferator-activated receptors (PPAR)in cell proliferation andor differentiation cell survivalautophagy senescence apoptosis and necrosis (see below 4-HNE as signaling molecule)

Characteristics of various lipid peroxidation products asbiomarkers have been reviewed on the basis of mechanismsand dynamics of their formation and metabolism and alsoon the methods of measurement with an emphasis on theadvantages and limitations [58]

23 Primary Lipid Peroxidation Product-Lipid Hydroperox-ides Hydroperoxides are produced during the propagationphase constituting the major primary product of lipid perox-idation process The hydroperoxide group may be attachedto various lipid structures for example free fatty acids tria-cylglycerols phospholipids and sterols Lipid hydroperoxidegeneration turnover and effector action in biological systemshave been reviewed [36] In contrast to free radical usuallyhighly reactive and chemically unstable at moderate reactionconditions such as low temperature and absence of metalions lipid hydroperoxides are relativelymore stable productsWe found that lipid hydroperoxides in serum could be usefulto predict the oxidative stress in tissues [59] and the levelsof oxidative stress including lipid peroxidation increasedthroughout the day [60] Once formed lipid hydroperoxidescan be target of different reduction reactions resultingin peroxidative damage inhibition or peroxidative damageinduction

Peroxidative Damage Inhibition Hydroperoxides maydecompose in vivo through two-electron reduction whichcan inhibit the peroxidative damage The enzymes mainlyresponsible for two-electron reduction of hydroperoxidesare selenium-dependent glutathione peroxidases (GPx)and selenoprotein P (SeP) GPxs are known to catalyze thereduction of H

2

O2

or organic hydroperoxides to water orthe corresponding alcohols respectively typically usingglutathione (GSH) as reductant Widely distributed in


OOH2 3

1

Rearrangement

Unsaturated lipid

Unsaturated lipid

Unsaturated lipid radical

Lipid peroxyl radical

Lipid hydroperoxide

Antioxidant4

O2

OO∙

R∙ H+

Figure 2 Lipid peroxidation process In Initiation prooxidants abstract the allylic hydrogen forming the carbon-centered lipid radical thecarbon radical tends to be stabilized by a molecular rearrangement to form a conjugated diene (step 1) In the propagation phase lipid radicalrapidly reacts with oxygen to form a lipid peroxy radical (step 2) which abstracts a hydrogen from another lipid molecule generating a newlipid radical and lipid hydroperoxide (step 3) In the termination reaction antioxidants donate a hydrogen atom to the lipid peroxy radicalspecies resulting in the formation of nonradical products (step 4)

mammalian tissues GPx can be found in the cytosol nucleiand mitochondria [61 62] The presence of selenocysteine(in the catalytic centre of glutathione peroxidases) as thecatalytic moiety was suggested to guarantee a fast reactionwith the hydroperoxide and a fast reducibility by GSH[61] SeP is the major selenoprotein in human plasma thatreduced phospholipid hydroperoxide using glutathione orthioredoxin as cosubstrate It protected plasma proteinsagainst peroxynitrite-induced oxidation and nitration orlow-density-lipoproteins (LDL) from peroxidation [62]

Peroxidative Damage Induction Hydroperoxides may alsodecompose in vivo through one-electron reduction and takepart in initiationpropagation steps [31 36 37] induce newlipid hydroperoxides and feed the lipid peroxidation processall these mechanisms can contribute to peroxidative damageinductionexpansion Lipid hydroperoxides can be convertedto oxygen radicals intermediates such as lipid peroxyl radical(LOO∙) andor alkoxyl (LO∙) by redox cycling of transitionmetal (M) resulting in lipid hydroperoxide decompositionand the oxidized or reduced formof thesesmetal respectively[63] The lipid peroxyl and alkoxyl radicals can attack otherlipids promoting the propagation of lipid peroxidation

LOOH +M119899 997888rarr LO∙ +OHminus +M119899+1 (1)

LOOH +M119899+1 997888rarr LOO∙ +H+ +M119899 (2)

Lipid hydroperoxides can also react with peroxynitrite (ashort-lived oxidant species that is a potent inducer of celldeath [64] and is generated in cells or tissues by the reactionof nitric oxide with superoxide radical) or hypochlorous

acid (a high reactive species produced enzymatically bymyeloperoxidase [65 66] which utilizes hydrogen peroxideto convert chloride to hypochlorous acid at sites of inflam-mation) yielding singlet molecular oxygen [67 68] Singletoxygen (molecular oxygen in its first excited singlet state 1Δ

119892

1O2

)1 can react with amino acid and proteins resulting inmultiple effects including oxidation of side-chains backbonefragmentation dimerizationaggregation unfolding or con-formational changes enzymatic inactivation and alterationsin cellular handling and turnover of proteins [69 70]

Major substrates for lipid peroxidation are polyunsatu-rated fatty acids (PUFAs) [31 36 37] which are a familyof lipids with two or more double bounds that can beclassified in omega-3 (n-3) and omega-6 (n-6) fatty acidsaccording to the location of the last double bond relative tothe terminalmethyl end of themoleculeThe predominant n-6 fatty acid is arachidonic acid (AA) which can be reduced (i)via enzymatic peroxidation to prostaglandins leukotrienesthromboxanes and other cyclooxygenase lipoxygenase orcytochrome P-450 derived products [4] or (ii) via nonen-zymatic peroxidation to MDA 4-HNE isoprostanes andother lipid peroxidation end-products (more stables and toxicthan hydroperoxides) through oxygen radical-dependentoxidative routes [49 71] The continued oxidation of fattyacid side-chains and released PUFAs and the fragmentationof peroxides to produce aldehydes eventually lead to lossof membrane integrity by alteration of its fluidity whichfinally triggers inactivation of membrane-bound proteinsContrary to radicals that attack biomolecules located lessthan a few nanometres from its site of generation [22] thelipid peroxidation-derived aldehydes can easily diffuse across


membranes and can covalently modify any protein in thecytoplasm and nucleus far from their site of origin [72]

24 Secondary Lipid Peroxidation ProductsMDA MDA is anend-product generated by decomposition of arachidonic acidand larger PUFAs [49] through enzymatic or nonenzymaticprocesses (Figure 3) MDA production by enzymatic pro-cesses is well known but its biological functions and its possi-ble dose-dependent dual role have not been studied althoughMDA is more chemically stable and membrane-permeablethanROS and less toxic than 4-HNE andmethylglyoxal (MG)[49] So far only few papers have reported that MDA mayact as signaling messenger and regulating gene expression(i) very recent research indicated that MDA acted as asignaling messenger and regulated islet glucose-stimulatedinsulin secretion (GSIS) mainly through Wnt pathway Themoderately high MDA levels (5 and 10 120583M) promoted isletGSIS elevated ATPADP ratio and cytosolic Ca2+ level andaffected the gene expression and proteinactivity productionof the key regulators of GSIS [73] (ii) in hepatic stellate cellsMDA induced collagen-gene expression by upregulatingspecificity protein-1 (Sp1) gene expression and Sp1 and Sp3protein levels [74] Both Sp1 and Sp3 can interact with andrecruit a large number of proteins including the transcrip-tion initiation complex histone modifying enzymes andchromatin remodeling complexes which strongly suggestthat Sp1 and Sp3 are important transcription factors in theremodeling chromatin and the regulation of gene expression[75] On the other hand MDA production by nonenzymaticprocesses remains poorly understood despite their potentialtherapeutic value because this MDA is believed to originateunder stress conditions and has high capability of reactionwith multiple biomolecules such as proteins or DNA thatleads to the formation of adducts [76ndash78] and excessiveMDAproduction have been associated with different pathologicalstates [79ndash85] (see Table 1) Identifying in vivoMDA produc-tion and its role in biology is important as indicated by theextensive literature on the compound (over 15 800 articles inthe PubMed database using the keyword ldquomalondialdehydelipid peroxidationrdquo in December 2013)

MDA Production by Enzymatic Processes MDA can begenerated in vivo as a side product by enzymatic processesduring the biosynthesis of thromboxane A

2

(Figure 3) [86ndash90] TXA

2

is a biologically active metabolite of arachidonicacid formed by the action of the thromboxane A2 synthaseon prostaglandin endoperoxide or prostaglandin H2 (PGH

2

)[4 91 92] PGH

2

previously is generated by the actions ofcyclooxygenases on AA [4 91 93]

MDA Production by Nonenzymatic Processes A mixture oflipid hydroperoxides is formed during lipid peroxidationprocess The peroxyl radical of the hydroperoxides with acis-double bond homoallylic to the peroxyl group permitstheir facile cyclization by intramolecular radical addition tothe double bond and the formation of a new radical Theintermediate free radicals formed after cyclization can cyclizeagain to form bicycle endoperoxides structurally relatedto prostaglandins and undergo cleavage to produce MDA

Through nonenzymatic oxygen radical-dependent reactionAA is the main precursor of bicyclic endoperoxide whichthen undergoes further reactions with or without the partic-ipation of other compounds to form MDA (Figure 3) [31 4994 95] However it should be possible that other eicosanoidsthat can also be generated by nonenzymatic oxygen radical-dependent reaction [96ndash99] may be precursor of bicyclicendoperoxide and MDA Recent review has addressed thepathways for the nonenzymatic formation of MDA underspecific conditions [100]

MDA Metabolism Once formed MDA can be enzymaticallymetabolized or can react on cellular and tissular proteins orDNA to form adducts resulting in biomolecular damagesEarly studies showed that a probable biochemical routefor MDA metabolism involves its oxidation by mitochon-drial aldehyde dehydrogenase followed by decarboxylationto produce acetaldehyde which is oxidized by aldehydedehydrogenase to acetate and further to CO

2

and H2

O(Figure 3) [49 101 102] On the other hand phosphoglucoseisomerase is probably responsible for metabolizing cytoplas-mic MDA to methylglyoxal (MG) and further to D-lactateby enzymes of the glyoxalase system by using GSH as acofactor [103] A portion of MDA is excreted in the urine asvarious enaminals (RNH-CHndashCH-CHO) such as N-epsilon-(2-propenal)lysine or N-2-(propenal) serine [49]

241 MDA Biomolecules Adducts As a bifunctional elec-trophile aldehyde MDA reactivity is pH-dependent whichexists as enolate ion (conjugate bases having a negativecharge on oxygen with adjacent CndashC double bond) with lowreactivity at physiological pH When pH decreases MDAexists as beta-hydroxyacrolein and its reactivity increases[49] MDArsquos high reactivity is mainly based on its elec-trophilicity making it strongly reactive toward nucleophilessuch as basic amino acid residues (ie lysine histidine orarginine) Initial reactions between MDA and free aminoacids or protein generate Schiff-base adducts [49 104 175]These adducts are also referred to as advanced lipid per-oxidation end-products (ALEs) Acetaldehyde (product ofMDAmetabolism) under oxidative stress and in the presenceof MDA further generates malondialdehyde acetaldehyde(MAA) adducts [157 176] MAA adducts are shown to behighly immunogenic [177ndash181]MDAadducts are biologicallyimportant because they can participate in secondary delete-rious reactions (eg crosslinking) by promoting intramolec-ular or intermolecular proteinDNA crosslinking that mayinduce profound alteration in the biochemical properties ofbiomolecules and accumulate during aging and in chronicdiseases [72 104 182 183] Important proteins that can bemodified by MDA adducts are as follows (i) eElongationfactor 2 (eEF2) catalyzes themovement of the ribosome alongthe mRNA in protein synthesis MDA adducts with eEF2could contribute to decline of protein synthesis secondary toLP increase (see belowmdashcumene hydroperoxide-induced lipidperoxidation) (ii) factor H (FH) is the main regulator of thealternative pathway in plasma that tightly controls the activa-tion of complement to prevent attack against host cells MDA


Table 1 Common pathological processes linked to MDA and 4-HNE

Pathological processes Aldehyde References

Alzheimerrsquos disease MDA4-HNE

[104ndash113][81 108 114ndash121]

Cancer MDA4-HNE

[109 122ndash130][72 126ndash128 131ndash136]

Cardiovascular diseases MDA4-HNE

[72 79 109 123 135 137ndash141][72 104 109 131 135 138 139 142ndash144]

Diabetes MDA4-HNE

[79 109 123 140 145ndash150][131 135 142 143 151ndash156]

Liver disease MDA4-HNE

[123 135 157ndash164][135 160ndash163 165ndash169]

Parkinsonrsquos disease MDA4-HNE

[81 108 114ndash121][72 114 131 135 142 170ndash174]

PUFAAA

Oxy radical Lipid hydroperoxide

Bicyclicendoperoxide

Monocyclicperoxide

HHTMDA

Malonicsemialdehyde

MDA-protein adductsMDA-DNA adducts

AcetaldehydeAcetateAcetylCoA

Biomolecular damagecell death

1

2

3 33

4

546

7

Cyclization

CO2 + H2O

O2

O2

+ H+

H+

H+

PUFA peroxide-radical∙

Radical∙

PUFA-radical∙

2O2

PGG2

PGH2

TXA2

Figure 3 MDA formation and metabolism MDA can be generated in vivo by decomposition of arachidonic acid (AA) and larger PUFAsas a side product by enzymatic processes during the biosynthesis of thromboxane A

2

(TXA2

) and 12-l-hydroxy-5810-heptadecatrienoic acid(HHT) (blue pathway) or through nonenzymatic processes by bicyclic endoperoxides produced during lipid peroxidation (red pathway)One formed MDA can be enzymatically metabolized (green pathway) Key enzymes involved in the formation and metabolism of MDAcyclooxygenases (1) prostacyclin hydroperoxidase (2) thromboxane synthase (3) aldehyde dehydrogenase (4) decarboxylase (5) acetylCoA synthase (6) and tricarboxylic acid cycle (7)


adducts with FH can block both the uptake ofMDA-modifiedproteins by macrophages and MDA-induced proinflamma-tory effects in vivo in mice [184] MDA adducts or MAAadducts can promote binding of complement (iii) anaphyla-toxin C3a (proinflammatory complement components) withoxidatively modified low-density lipoproteins (Ox-LDL) andcontributes to inflammatory processes involving activationof the complement system in atherosclerosis [185] and (iv)protein kinase C (PKC) is known to play a major role inintracellular signal transduction affecting such processes asproliferation differentiation migration inflammation andcytoskeletal organization BSA-MAA induces the activationof a specific isoform of PKC PKC-120572 in hepatic stellate cells(HSCs) and induces the increased secretion of urokinase-type plasminogen activator a key component of the plasmin-generating system thereby contributing to the progressionof hepatic fibrosis [186] A recent review shows a list of upto thirty-three proteins known to be modified by MDA andincluding enzymatic proteins carrier proteins cytoskeletalproteins and mitochondrial and antioxidant proteins [76]

It has also been proposed that MDA could react phys-iologically with several nucleosides (deoxy-guanosine andcytidine) to form adducts to deoxyguanosine and deoxya-denosine and the major product resulting is a pyrimidop-urinone called pyrimido[12-a]purin-10(3H-)one (M1G orM1dG) [122 123 187 188] MDA is an important contributorto DNA damage and mutation [122 124] The main route forrepair of M1dG residues in genomic DNA appears to be thenucleotide excision repair (NER) pathway [188 189] In theabsence of repair MDA-DNA adducts may lead to mutations(point and frameshift) [124] strand breaks [122 190] cellcycle arrest [191] and induction of apoptosis [192] M1dG isoxidized to 6-oxo-M1dG in rats and that xanthine oxidase(XO) and aldehyde oxidase (AO) are the likely enzymesresponsible [193] This MDA-induced DNA alteration maycontribute significantly to cancer and other genetic diseasesHypermethylated in cancer 1 (HIC1) is a tumor suppressorgene that cooperates with p53 to suppress cancer develop-ment New funding has shown that highest HIC1methylationlevels in tobacco smokers were significantly correlated withoxidative DNA adducts M1dG [125] Research also suggeststhat persistent M1dG adducts in mitochondrial DNA hinderthe transcription ofmitochondrial genes [194] Dietary intakeof certain antioxidants such as vitamins was associated withreduced levels of markers of DNA oxidation (M1dG and 8-oxodG) measured in peripheral white blood cells of healthysubjects which could contribute to the protective role ofvitamins on cancer risk [195]

25 Secondary Lipid Peroxidation Products 4-HNE 4-Hydroxynonenal (4-HNE) 120572 120573-unsaturated electrophiliccompounds is the major type of 4-hydroxyalkenals end-product generated by decomposition of arachidonic acid andlarger PUFAs through enzymatic or nonenzymatic processes[49] 4-HNE is an extraordinarily reactive compound con-taining three functional groups (i) C=C double bond thatcan be target to Michael additions to thiol reduction orepoxidation (ii) carbonyl group which can yield acetalthio

acetal or can be target to Schiff-base formation oxidation orreduction and (iii) hydroxyl group which can be oxidized toa ketone [56]

4-HNE is the most intensively studied lipid peroxidationend-product in relation not only to its physiological andprotective function as signaling molecule stimulating geneexpression but also to its cytotoxic role inhibiting geneexpression and promoting the development and progressionof different pathological states In the last three yearsexcellent reviews have been published summarizing bothsignaling and cytotoxic effects of this molecule in biology forexample overview of mechanisms of 4-HNE formation andmost common methods for detecting and analyzing 4-HNEand its protein adducts [196] Review focuses on membraneproteins affected by lipid peroxidation-derived aldehydesunder physiological and pathological conditions [131]Jaganjac andCo-workers have described the role of 4-HNE assecond messengers of free radicals that act both as signalingmolecules and as cytotoxic products of lipid peroxidationinvolvement in the pathogenesis of diabetes mellitus (DM)[151] Chapple and Co-workers summarized the productionmetabolism and consequences of 4-HNE synthesis withinvascular endothelial smooth muscle cells and targetedsignaling within vasculature [142] Review focuses on the roleof 4-HNE and Ox-PLs affecting cell signaling pathways andendothelial barrier dysfunction through modulation of theactivities of proteinsenzymes byMichael adducts formationenhancing the level of protein tyrosine phosphorylation ofthe target proteins and by reorganization of cytoskeletalfocal adhesion and adherens junction proteins [197] Anoverview of molecular mechanisms responsible for theoverall chemopreventive effects of sulforaphane (SFN)focusing on the role of 4-HNE in these mechanismswhich may also contribute to its selective cytotoxicity tocancer cells [198] Perluigi and Co-workers summarized therole of lipid peroxidation particularly of 4-HNE-inducedprotein modification in neurodegenerative diseasesIn this review the authors also discuss the hypothesisthat altered energy metabolism reduced antioxidantdefense and mitochondrial dysfunction are characteristichallmarks of neurodegenerative [170] Zimniak describedthe effects of 4-HNE and other endogenous electrophiles onlongevity and its possible molecular mechanisms The roleof electrophiles is discussed both as destabilizing factorsand as signals that induce protective responses [199] Reedshowed the relationship between lipid peroxidation4-HNE and neurodegenerative diseases It also demonstrateshow findings in current research support the commonthemes of altered energy metabolism and mitochondrialdysfunction in neurodegenerative disorders [171] Fritzand Petersen summarized the generation of reactivealdehydes via lipid peroxidation resulting in proteincarbonylation and pathophysiologic factors associated with4-HNE-protein modification Additionally an overviewof in vitro and in vivo model systems used to study thephysiologic impact of protein carbonylation and an updateof the methods commonly used in characterizing proteinmodification by reactive aldehydes [200] Butterfield and Co-workers showed that several important irreversible protein


modifications including protein nitration and 4-HNEmodification both which have been extensively investigatedin research on the progression of Alzheimerrsquos disease (AD)[201] Balogh and Atkins described the cellular effectsof 4-HNE followed by a review of its GST-catalyzeddetoxification with an emphasis on the structural attributesthat play an important role in the interactions with alpha-class GSTs Additionally a summary of the literature thatexamines the interplay between GSTs and 4-HNE in modelsystems relevant to oxidative stress is also discussed todemonstrate the magnitude of importance of GSTs in theoverall detoxification scheme [202] Like MDA 4-HNE hashigh capability of reaction with multiple biomolecules suchas proteins or DNA that lead to the formation of adducts[49]

4-HNE Production by Enzymatic Processes 4-HNE is alipid peroxidation end-product of enzymatic transforma-tion of n-6 PUFAs (AA linoleic acid and other) by 15-lipoxygenases (15-LOX) Two different 15-LOX exist (i)15-LOX-1 (reticulocyte type) expressed in reticulocyteseosinophils and macrophages (ii) and 15-LOX-2 (epidermistype) expressed in skin cornea prostate lung and esophagus[203ndash205] Mice do not express 15-LOX and only expressthe leukocyte-derived 12-LOX In plant enzymatic routeto 4-HNE includes lipoxygenase (LOX) -hydroperoxidelyase (HPL) alkenal oxygenase (AKO) and peroxygenases(Figure 4) [206] The main precursors of 4-HNE in humanare 13-hydroperoxyoctadecadienoic acid (13-HPODE) pro-duced by the oxidation of linoleic acid by 15-LOX-1 [207] and15- hydroperoxyeicosatetraenoic acids (15-HPETE) producedby the oxidation of AA by 15-LOX-2 [208]These compoundsare short lived and are catabolised into various familiesof more stable compounds such as 15-HETEs lipoxinsand leukotrienes [4] 15-HPETE is associated with anti-inflammatory and proapoptotic functions (the release ofcytochrome c activation of caspase-3 and 8 PARP and Bidcleavage) and DNA fragmentation [209 210]

4-HNE Production by Nonenzymatic Processes 4-HNE canbe formed through several nonenzymatic oxygen radical-dependent routes involving the formation of hydroperoxidesalkoxyl radicals epoxides and fatty acyl crosslinking reac-tions Spickett C [196] recently reviewed the mechanisms offormation of 4-HNE during lipid peroxidation and showedthat the main processes leading to 4-HNE are likely beta-cleavage reaction of lipid alkoxy-radicals which can besummarized into five generic mechanisms (i) reduction ofthe hydroperoxide to a lipid alkoxy radical by transitionmetalions such as Fe2+ followed by b-scission (ii) protonation ofthe lipid hydroperoxide yields an acidified lipid hydroperox-ide that undergoes Hock rearrangement of a CndashC to CndashObond followed by hydrolysis and Hock cleavage (iii) the lipidperoxyl radical of the hydroperoxides permits their facilecyclization to dioxetane and ending with dioxetane cleavage(iv) free radical attack to 120596-6 PUFA on bis-allyl site yieldinga free radical intermediate that further reacts with molecularoxygen to generate hydroperoxide derivatives such as 13-HPODE or 15-HPETEThe abstraction of an allylic hydrogen

LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA

Figure 4 Enzymatic production of 4-HNE and metabolism Inplant enzymatic route to 4-HNE includes lipoxygenase (LOX)-hydroperoxide lyase (HPL) alkenal oxygenase (AKO) and per-oxygenases 4-HNE metabolism may lead to the formation ofcorresponding alcohol 14-dihydroxy-2-nonene (DHN) corre-sponding acid 4-hydroxy-2-nonenoic acid (HNA) and HNEndashglutathione conjugate products 4-HNE conjugation with glu-tathione s-transferase (GSH) produce glutathionyl-HNE (GS-HNE)followed by NADH-dependent alcohol dehydrogenase (ADH-)catalysed reduction to glutathionyl-DNH (GS-DNH) andor alde-hyde dehydrogenase (ALDH-)catalysed oxidation to glutathionyl-HNA (GS-HNA) 4-HNE is metabolized by ALDH yielding HNAwhich is metabolized by cytochrome P450 (CYP) to form 9-hydroxy-HNA (9-OH-HNA) 4-HNE may be also metabolized byADH to produce DNH

of their structure produce another radical intermediate thatafter oxygenation step forms the corresponding dihydroper-oxyde derivative (unstable) which after Hock rearrange-ment and cleavage produces 4-hydroperoxy-2E-nonenal (4S-HPNE) immediate precursor of HNE and (v) the oxida-tion products generated after reaction of linoleate-derivedhydroperoxy epoxide (13-Hp-Epo-Acid) with Fe+2 yields analkolxyl radical which undergo to di-epoxy-carbinyl radicaland after beta-scission yield different aldehydes compoundsincluding 4-HNE (Figure 5)

Once formed 4-HNE and depending of cell type andcellular metabolic circumstances can promote cell survival ordeath Cells expressing differentiated functions representativefor the in vivo situation react more sensitively to 4-HNE thancell linesThe different response with respect to the endpointsof genotoxicity probably depends on the different metabo-lizing capacities and thus the action of different metabolitesof 4-HNE [211] 4-HNE can be enzymatically metabolizedat physiological level and cells can survive 4-HNE can playan important role as signaling molecule stimulating geneexpression (mainly Nrf2) with protective functions that canenhance cellular antioxidant capacity and exert adaptiveresponse when 4-HNE level is low under this circumstances


PUFAlipoic acid

9 10 dioxetane

4-HPNE

Hydroperoxyl dioxetaneCyclization

Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+

Radical∙Radical∙

13-Lipid radical∙

13-Peroxyl radical ∙9-Peroxyl radical ∙

13-Hydroperoxyl radical ∙

O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙

9-Hydroperoxyl radical∙

9-Alkoxyl radical∙


4-HNE

120573-Scission

Figure 5 Nonenzymatic 4-HNE production Initial abstraction of bisallylic hydrogen of lipoic acid (LA) produces fatty radicals 4-HNEformation starting with 9- and 13-hydroperoxyoctadecadienoate (HPODE) (red and blue pathways resp) 4-HNE is generated by beta-scission of a hydroxyalkoxy radical that is produced after cyclization of alkoxy radical in the presence of transition metal ions and twomolecules of oxygen this reaction involves hydrogen abstraction (1) Peroxy radical cyclizes to form a dioxetane which is oxygenatedto peroxy-dioxetane that is fragmented and after two hydrogen abstractions produce 4-HNE (2) Hydroperoxyl radical is oxygenated todioxetane that is further fragmented to produce 4-hydroperoxy-2E-nonenal (4-HPNE) an immediate precursor of 4-HNE (3) Bicyclicendoperoxides react with reduced form of transition metal such as iron (Fe2+) to produce alkoxyl radicals which after reaction with oxygen(O2

) hydrogen abstraction (H+) and fragmentation produce 4-HNE (4) Alkoxyl radical after cyclization oxygenation hydrogen abstractionoxidation of transitionmetal hydrolysis and rearrangement yields 4-HNE (5)With arachidonic acid 11- and 15- hydroperoxyeicosatetraenoicacids (HPETE) are the precursors to form 4-HNE via the analogous mechanisms

cells can survive 4-HNE can promote organelle and proteindamage leading to induction of autophagy senescence or cellcycle arrest at 4-HNEmedium level and cells can subsist andfinally 4-HNE induces apoptosis or necrosis programmedcell death at 4-HNE high or very high level respectivelyand cells die These processes eventually lead to molecularcell damage which may facilitate development of variouspathological states High levels of 4-HNE can also react withproteins andor DNA to form adducts resulting in a varietyof cytotoxic and genotoxic consequences (Figure 6)

4-HNE Metabolism The main goal of the rapid intracellularmetabolism of 4-HNE in mammalian cells is to protectproteins from modification by aldehydic lipid peroxida-tion products [212] The biochemical routes of 4-HNEmetabolism that lead to the formation of correspondingalcohol 14-dihydroxy-2-nonene (DHN) corresponding acid4-hydroxy-2-nonenoic acid (HNA) and HNE-glutathione

conjugate products can be summarized according to stresslevels (i) under physiological or low stress levels the major4-HNE detoxification step is conjugation with GSH to yieldglutathionyl-HNE (GS-HNE) or glutathionyl-lactone (GS-)lactone (cyclic ester 4-HNE- form) followed by NADH-dependent alcohol dehydrogenase (ADH-)catalysed reduc-tion to glutathionyl-DNH (GS-DNH) andor aldehyde dehy-drogenase (ALDH-)catalysed oxidation to glutathionyl-HNA(GS-HNA) (ii) at moderate stress levels 4-HNE undergoesaldehyde dehydrogenase (ALDH-)catalysed oxidation yield-ing HNA that may be further metabolized in mitochondriathrough beta-oxidation by cytochrome P450 to form 9-hydroxy-HNA and (iii) at high stress levels 4-HNE ismetabolized byADH (that belongs to the aldo-keto reductase(AKR) superfamily) to produce DNH [131 196 202 212 213](Figure 4) By disrupting the Gsta4 gene that encodes thealpha class glutathione s-transferase (GST) isozyme GSTA4-4 in mice showed that GSTA4-4 plays a major role in


Cell signalingand response

to stress

Cellularantioxidantinduction

Physiologicallevels

Cell signalingprotein damage

Autophagysenescence orcell cycle arrest

4-HNE ismetabolized

Adducts andapoptosis

Development of pathological

states

Irreversible cell injurydamage

Programmednecrosis cell

death

Lowlevels

Mediumlevels

Highlevels

Very highlevels

Cell subsist Cell dieCell survive Cell survive

4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE 4-HNE 4-HNE

Cell die

Figure 6 4-HNE promotes cell survival or induces cell death Depending on cell type damagerepair capacities and cellular metaboliccircumstances 4-HNE can promote cell survival or induce cell death 4-HNE at physiological levels is enzymatically metabolized and at lowlevels plays an important role as signaling molecule stimulating gene expression enhance cellular antioxidant capacity and exert adaptiveresponse at medium levels organelle and protein damage lead to induction of autophagy senescence or cell cycle arrest and at high or veryhigh levels promote adducts formation and apoptosis or necrosis cell death respectively

protecting cells from the toxic effects of oxidant chemicals byattenuating the accumulation of 4-HNE [214] Overexpres-sion and inhibition of ALDH activity reduce and increaserespectively the 4-HNE toxicity and 4-HNE-protein adductslevels in cell culture [215 216]

251 4-HNE as Signaling Molecule At moderate concentra-tion when the basal level of antioxidant enzymes cannotbe sufficient to neutralize 4-HNE cells can survive due to4-HNE may regulate several transcription factors sensibleto stress such as nuclear factor erythroid 2-related factor 2(Nrf2) activating protein-1 (AP-1) NF-120581B and peroxisome-proliferator-activated receptors (PPAR) It also activatesstress response pathways such as mitogen-activated proteinkinases (MAPK) EGFRAkt pathways and protein kinaseC Different labs demonstrated the 4-HNE-dependent induc-tion of Nrf2 a primary sensor and oxidative stress regulator[217ndash221] Also administration of the Nrf2-ARE activatorsprotect from 4-HNE toxicity [222] Under physiological con-ditions Nrf2 is sequestered in the cytoplasm by the repressorprotein Keap1 but in response to oxidant stimuli Nrf2 isactivated and translocated into the nucleus wheremediate thetranscription of antioxidantcytoprotective genes by bindingto the antioxidant-response element (ARE) within DNA[223] The Nrf2-ARE pathway has essential role in differentpathological states such as neurodegenerative diseases [223]cancer [224] diabetes [225] and infectious disease [226]Themain genes regulated by 4-HNE- inducedNrf2-ARE pathwayare as follows (i) HO-1 an antioxidant protein that catalyzesthe degradation of heme to biliverdin which is then degradedto bilirubin both biliverdin and bilirubin have antioxidantproperties [227] 4-HNE can upregulate HO-1 [217 220 221228ndash230] (ii) thioredoxin (Trx) and thioredoxin reductase(TrxR) Trx is a small (13 kDa) antioxidant ubiquitous protein

with two redox-active cysteine residues (-Cys-Gly-Pro-Cys-)in its active center oxidized Trx is reduced back to theactive form of Trx by Trx reductase (TrxR) in the presenceof NADPH [231] 4-HNE can upregulate TrxTrxR [220221 232] (iii) glutamate cystein ligase (GCL) is a majordeterminant enzyme inGSH synthesis [233 234] 4-HNE canupregulate GCL [235ndash239]

Involvement of AP-1 transcription factor in 4-HNE-induced cell signaling has been demonstrated by severalstudies which showed an AP-1 upregulation by 4-HNE [240ndash243] Activation of AP-1 binding may lead to the 4-HNE-induced increase in GSH content [239] AP-1 is a dimer con-sisting of basic region-leucine zipper proteins from the Junand Fos subfamilies AP-1 transcription factors control cellproliferation survival and death Growth factors cytokinescellular stress and many other stimuli activate AP-1 [244245]

NF-120581B is a dimeric transcription factor that regulatesdiverse biological processes including immune responsesinflammation cell proliferation and apoptosis The NF-120581B protein complex is retained in an inactive state in thecytoplasm by binding to inhibitory proteins I120581Bs family[246] Various cellular stimuli such as oxidative stress I120581Bsare phosphorylated making them susceptible to degradationby the ubiquitin-proteasome system This results in nucleartranslocation of NF-120581B complex where it can bind to variouspromoter areas of its target genes and induce gene tran-scription of the corresponding genes [246 247] most ofwhich are implicated in the regulation of inflammation 4-HNE can activate or inhibit NF-120581B depending on the typeof cells used For example 4-HNE inhibited the activity ofNF-120581B in hepatocytes [165] cortical neurons [248] ARPE-19 human retinal pigment epithelial cells [249] Kupffercells [250] human aortic endothelial cells [251] humancolorectal carcinoma and lung carcinoma cell [252] On the


contrary 4-HNE induced activity of NF-120581B in macrophages[253] vascular smooth muscle cells [254] PC12 cells [255]optic nerve head astrocytes [256] human osteoarthriticchondrocytes [257] human fibroblasts [258] and humanmonocytic lineage cells [259]

PPARs comprise three subtypes (PPAR120572 120573120575 and 120574)to form a nuclear receptor superfamily PPARs act as keytranscriptional regulators of lipidmetabolismmitochondrialbiogenesis and antioxidant defense [260 261] PPARs inter-actionmodulation with 4-HNE has been reviewed [262]4-HNE increased PPAR-120574 gene expression and acceleratedadiponectin protein degradation in adipocytes [263] expres-sion of PPAR-120574 was induced in HL-60 and U937 cells by4-HNE treatment [264] whereas in the colon cancer cell(CaCo-2) PPAR120574 protein expression was not induced after 4-HNE treatment [265] 4-HNE increased PPAR1205742 expressionin C2C12 cells [266] PPAR-120573120575 is activated by 4-HNE in 3T3-L1 preadipocytes cells [267] 4-HNE activates PPAR-120575 andamplifies insulin secretion in INS-1E 120573-cells [152]

MAP kinases family can be activated in response todiverse stimuli such as oxidative stress lipopolysaccharidesinflammatory cytokines growth factors or endoplasmicreticulum (ER) stress and are involved in several cellu-lar responses like cell proliferation andor differentiationinflammation proteasomal-mediated protein degradationand apoptosis Members of the major mitogen-activatedprotein kinase (MAPK) subfamilies are the extracellularsignal-regulated kinase (ERK) p38 and Jun N-terminalkinase (JNK) subfamilies The mechanism by which MAPKsignaling cascades are activated by 4-HNE is not well knownFor example activation of different MAPK under variousstimuli can affect both apoptotic and prosurvival signalingIn corneal epithelial cells 4-HNE caused a time-dependentinduction of HO-1 mRNA and protein via modificationand activation of Erk12 JNK and p38 MAP kinases aswell as phosphoinositide-3-kinase (PI3)Akt Inhibition ofp38 blocked 4-HNE-induced HO-1 expression inhibition ofErk12 and to a lesser extent JNK and PI3KAkt suppressed4-HNE-induced HO-1 [268] 4-HNE also stimulated Erk12JNK p38 and PI3 kinase in keratinocyte and the inhibitorsof these enzymes suppressed 4-HNE-induced expression ofHO-1 [269] In PC12 cells 4-HNE treatment induced ERKJNK and p38 MAPK activation as well as induced theexpression of HO-1 Addition of p38MAPK specific inhibitorSB203580 attenuated HO-1 upregulation these results indi-cate that 4-HNE-induced transient p38 MAPK activationmay serve as an upstream negative regulator of ER stressand confer adaptive cytoprotection against 4-HNE-mediatedcell injury [228] In rat liver epithelial RL34 cells 4-HNEupregulates the cyclooxygenase-2 (COX-2 which plays a keyrole in conversion of free arachidonic acid to PGs) expressionby the stabilization of COX-2 mRNA via activation of thep38 MAPK pathway [270] In human hepatic stellate cells(hHSC) 4-HNE forms adducts with JNK and this eventleads to JNK nuclear translocation and activation as wellas to c-jun and AP-1 induction [271] In human bronchialepithelial cells 4-HNE downmodulates the protein-tyrosinephosphatase SH2 domain containing phosphatase-1 (SHP-1)which negatively regulates JNK activity [272]We can also see

the protective effects of MAPK activation via GSH inductionbecause the activation of the ERK pathway is involved inGCL(the rate-limiting enzyme in de novo glutathione (GSH)synthesis) regulation in rat cells [273] while the JNKpathwaysappear to be involved in human HBE-1 cells [274]

In human monocytes 4-HNE was shown to significantlyinhibit p38 and ERK activity which resulted in inhibition ofTNF and interleukin-1beta production in response to LPSThe data suggest that 4-HNE at nontoxic concentrations hasanti-inflammatory properties [275] In human osteoarthriticosteoblasts 4-HNE also showed a significant (approximately70) decrease of TNF-120572-induced IL-6 mRNA expression viathe NF-120581B signaling pathway However only p38 MAPK andJNK12 were activated but not ERK12 [276] while 4-HNEalso inducedCOX-2 expression and prostaglandin E2 (PGE2)release [257 276]

On the other hand 4-HNE mediated depletion of intra-cellular thiols protein tyrosine phosphorylation MAPK(JNK ERK and p38) activation and modulates integrinresulting in reorganization of cytoskeletal focal adhesionproteins and barrier dysfunction in lung microvascularendothelial cells [277] Results suggest that activation andphosphorylation of MAP kinases (JNK ERK and p38) playan important role in 4-HNE mediated toxicity and celldeath in mouse embryonic fibroblasts (MEF) and absenceof GSTA4ndash4 potentiates the cytotoxic effects of 4-HNE Theincrease of apoptosis in Gsta4 null MEF by 4-HNE was asso-ciated with the enhanced accumulation of 4-HNE-proteinadducts DNA damage and the activation of caspases-3-8 and -9 [214] 4-HNE upregulates and phosphorylatescytosolic phospholipase A-2 (cPLA-2) in cultured microglialcell line (Ra2) via the ERK and p38 MAPK pathways [278]cPLA is a proinflammatory enzyme that stimulateAA- releaseby hydrolyzes glycerophospholipids with AA in the sn-2position

Matrix metalloproteinases (MMPs) constitute a largegroup of endoproteases that are not only able to cleave allprotein components of the extracellular matrix but also toactivate or inactivate many other signaling molecules suchas receptors adhesion molecules and growth factors [279]4-HNE induced MMP-9 production in macrophages [280]and MMP-2 in vascular smooth muscle cells (VSMC) [281]via activation of ERK and p38MAPKpathways consequentlyleading to plaque instability in atherosclerosis 4-HNE alsoenhances MMP-2 production in VSMC via mitochondrialROS-mediated activation of the AktNF-kappaB signalingpathways [254] In osteoarthritic (OA) synovial cells 4-HNEinduced MMP-13 mainly through activation of p38 MAPK[282]

Akt (aka protein kinase B or PKB) comprises three closelyrelated isoforms Akt1 Akt2 and Akt3 (or PKB120572120573120574 resp)which play a role in the regulation of cell proliferationsurvival and metabolism Dysregulation of Akt leads todiseases such as cancer diabetes and cardiovascular andneurological diseases [283] Under conditions of enhancedoxidative stress a major cellular response is the activa-tion of the Akt pathway that involves the oxidation andsubsequent inactivation of PTEN (phosphatase and tensinhomolog deleted on chromosome 10) a tumor suppressor


and primary regulator of Akt [284] Recent studies havealso demonstrated that activation of PI3 KAkt signaling by4-HNE occurs via modification and inhibition of PTENa regulatory protein that suppresses Akt2 activity whichis selectively phosphorylated by 4-HNE in both cellularhuman hepatocellular carcinoma cell line (HepG2) [285]and animal models (ethanol-fed mice) [286] In HepG2cells 4-HNE inhibits H

2

O2

-mediated activation of the Aktpathway in leading to phosphorylation of Akt1 but not Akt2decreased cell proliferation and decreased expression ofcyclin D1 [287] In retinal pigment epithelial (RPE) cells atlower concentrations 4-HNE triggered phosphorylation ofepidermal growth factor receptor (EGFR) and activation of itsdownstream signaling components ERK12 and Akt this ledto protective mechanism against oxidative stress [288] Akt-induced activity by 4-HNE promotes cell survival throughinduction of HO-1 mRNA and protein in corneal epithelialcells [268] and in keratinocyte [269] The inhibitors of Aktsuppressed 4-HNE-induced expression of HO-1

Protein kinases C (PKCs) are a family of multifunctionalenzymes that play crucial roles in the transduction of manycellular signals such as control of cell proliferation survivaland transformation by phosphorylating various targets ThePKC family consists of three different groups conventional(120572 1205731 1205732 and 120574) novel (120575 120576 120578 and 120579) and atypical (120577and 120582120591) Conventional and novel PKC isoforms are lipid-sensitive enzymes and calcium-dependent and are usuallyactivated by growth factors through stimulation of phos-pholipase C (PLC) which hydrolyzes phosphatidylinositol-45-bisphosphate (PIP2) to generate inositol triphosphate(IP3) and DAG [6 289] Cells can express more than onePKC isoform and individual PKCs can mediate differentbiological processes For example in human promyelo-cytic leukemia (HL-60) cells [290ndash292] and rat neutrophils[293] 4-HNE induced a significant increase of PLC activitywhich should result in an increased production of IP3 andDAG known to stimulate PKC [289] Phagocytes suchas granulocytes and monocytesmacrophages which engulfmicrobial intruders and effectively kill and eradicate theforeign bodies contain a membrane-associated NADPHoxidase that produces superoxide leading to other ROS withmicrobicidal tumoricidal and inflammatory activities [294]In RAW 2647 mouse macrophage cells 4-HNE exhibiteda concentration-dependent inhibition of ROS by adductionto PKC a protein vital in the assembly and activation ofNADPH oxidase [295] In rat hepatocyte PKC- isoformsactivity is differentially regulated by concentrations 4-HNEFor example PKC-120572 activity was decreased in a dose-dependentmanner by all concentrations of 4-HNE while lowconcentrations of 4-HNE increased PKC 120573I and to a muchgreater extent PKC120573II activities By contrast theywere unaf-fected or even inhibited by higher concentrations of 4-HNEThis PKC-dependent- 4-HNE regulation could be involvedin the traffic of secretory glycoproteins [296] In NT2 neu-rons low 4-HNE concentrations (similar to concentrationsdetected in AD brain tissue) induced a 2ndash6 fold increaseof intracellular amyloid 120573-protein (A120573) production thatwas concomitant with selective activation of 120573I and 120573IIPKC isoforms [297 298] In macrophages a marked and

early upregulation of monocyte chemoattractant protein 1(MCP-1) release occurs in response to low 4-HNE concen-trations most likely through of the increase in the activityof PKC-120573I and 120573II classic isoforms while the activation ofPKC-120575 appeared to be involved in LPS-stimulated cells [299]Treatment of macrophages with 4-HNE cell-permeableesters of glutathionyl-4-hydroxynonenal (GS-HNE) andglutathionyl-14-dihydroxynonane (GS-DHN) activated NF-120581B and PLCPKC Aldolase reductase catalyzes the reductionof GS-HNE to GS-DHN AR inhibitionablation preventedPLC PKC and IKKalphabeta and NF-120581B activation causedby 4-HNE and GS-HNE but not by GS-DHN suggests anovel role for a reduced glutathione-lipid aldehyde conjugate(such asGS-DHN) as an obligatorymediator of ROS-inducedcytotoxicity [300]

252 Effect of 4-HNE on Autophagy One of the most impor-tant processes for maintaining normal metabolic and redoxsignaling through degradation of damaged proteins andorganelles is autophagy-lysosomal pathway [301] 4-HNEcan promote protein-adducts leading to protein damage andto induction of autophagy-lysosomal pathway [302] a pro-cess that is increased by treatmentwith an autophagy stimula-tor rapamycin If autophagy is blocked with a PI3 K inhibitor3-methyladenine apoptotic cell death occurs [301 302] Sev-eral mechanisms by which 4-HNE induces autophagy havebeen reported For example 4-HNE promotes the formationof protein adducts that accumulate in the endoplasmicreticulum (ER) and led to autophagy in rat aortic smoothmuscle cells through selective activation of the PKR-like ERkinase (PERK) pathway accompanied by JNK activation theupregulation of the HO-1 increased microtubule-associatedprotein 1 light chain 3 (LC3) formation and maintenance ofcell viability under conditions of excessive 4-HNE-proteinadducts accumulation [303] In differentiated SH-SY5Y neu-roblastoma cells glucose-dependent autophagy serves as aprotective mechanism in response to 4-HNE because low4-HNE-concentrations increased autophagy and inducedconcentration dependent CASP3caspase-3 activation andcell death Additionally inhibition of glucose metabolism by2-deoxyglucose and glycolysis by koningic acid a GAPDHinhibitor led to autophagy inhibition and increased CASP3activation and cell death [304] On the contrary phagocy-tosis of 4-HNE- and MDA-modified photoreceptor outersegments (POS) induced a marked reduction of autophagicactivity by 40 in retinal pigment epithelium (RPE) cellswhich may contribute to RPE cell dysfunction and degener-ation In contrast unmodified POS had no significant effecton autophagy [305]

253 Effect of 4-HNE on Senescence Cellular senescencedefined as arrest during the cell cycle (G0) is involvedin the complex process of the biological aging of tissuesorgans and organisms Senescence is driven by many factorsincluding oxidative stress the DNA damagerepair responseinflammation mitogenic signals and telomere shorteningTelomeres are considered a ldquobiological clockrdquo of the celland are shortened by each cell division until a critical


length is reached and dysfunction ensues Rapid telomereshortening may indicate a very high cellular activity DNA-repair pathways are then recruited and cells enter senescencelosing their capacity to proliferate In addition to cell divisionfactors causing telomere shortening include DNA damageinflammation and oxidative stress [306] Activation of aDNA damage response including formation of DNA damagefoci containing activated H2AX (120574-histone 2AX) at eitheruncapped telomeres or persistent DNA strand breaks is themajor trigger of cell senescence 120574H2AX is a sensitive markerof DNA damage particularly induction of DNA double-strand breaks [307] The length of telomeres depends on thetelomerase activity and the catalytic subunit of telomerase(hTERT) which is strongly upregulated in most human can-cers [308] and the major consequence of the reactivation oftelomerase activity is that tumor cells escape from senescenceThe expression of c-myc (an activator) mad-1 (a repressor)and sp-1 (an activatorrepressor) which have been shownto activate hTERT transcription The formation of 4-HNE-proteins adducts in general increased as a function of age[309] Quantitative evaluation showed that the majority ofsenescent hepatocytes (as measured by 120574-H2AX) were alsopositive for 4-HNE [310 311] 4-HNE can induce prematuresenescence by a direct suppression of telomerase activityaffecting the expression of hTERT In endothelial cells (EC)isolated and cultured from arterial segments of patients withsevere coronary artery disease chronic treatment with anantioxidant (that significantly decreased the levels of lipidperoxidation that is 4-HNE expression) N-acetyl-cysteinNAC significantly delayed cellular senescence via decrease ofDNA damage marker (120574H2AX) decrease of nuclear p53 andincrease in hTERT activity [312] In three human leukemiccell lines (HL-60 U937 and ML-1) [313] and in coloncancer cells (Caco-2 and HT-29) [314] telomerase activityand hTERT expression were downregulated by 4-HNE as aconsequence of downregulation of c-myc mRNA expressionand c-Myc DNA binding activity as well as upregulation ofmad-1 mRNA expression and Mad-1 DNA binding activityOn the other hand 4-HNE may induce cellular senescencethrough activation of critical cell cycle sentinels that mediatethis process such as the tumor suppressor proteins p53(see below) which is well known to play a central role insenescence [315ndash320] p53 protects cells of oxidative stressand promotes DNA repair However when in the cells theextent of damage overwhelms repair capacities p53 inducescell death [315ndash319] All these data thus confirmed a cell-specific association between senescence and 4-HNE

254 Effect of 4-HNE on Cell Cycle and Proliferation In cellcycle the transition of different phases is driven by severalphase-specific cyclin-CDK (cyclin-dependent kinase) com-plexes which previously have been activated In response tomitogens cyclin D is activated and phosphorylate retinoblas-toma protein (RB) which leads to activation of E2F proteinsand the expression of E2F-responsive genes inducing cells toreenter the cell cycle fromquiescence calledG0 toG1Activa-tion of E2F leads to the transcription of cyclin E for transitionfrom G1 to S phase Subsequent expression of cyclin A leads

to transition of S to G2 and cyclin B leads G2 to M phases[321 322] The promitotic factor Cdc25 stimulates cell cycleprogression through the activation of cyclin A-Cdk1 cyclinB-Cdk1 and cyclin E-Cdk2 for entry intoM phase by remov-ing the inhibitory phosphorylation on Cdk1 and Cdk2 Onthe contrary the anti-mitotic factor (p21 p27 p57) inhibit cellcycle progression through inhibition of cyclin AndashCdk1 cyclinBndashCdk1 cyclin EndashCdk2 and cyclin DndashCdk46 [321ndash323] Inresponse to 4-HNE the expression of key components of cellcycle can be modulated and cells are arrested at G1 or G2Several studies showed that in general 4-HNE may inducecell cycle arrest in malignant cell and inhibition or decreaseof cell proliferation For example treatment of HL-60 cellswith 4-HNE (1120583M) causes a p53-independent increase ofp21 expression RB dephosphorylation progressive reductionin the amount of free E2F bound to DNA and a relativeincrease in E2F complexes at higher molecular weights withrepressive activity decrease of E2F complexes [324] anddecrease of cyclinD1 cyclinD2 and cyclin A [325] In humanerythroleukemia cells (K562) 4-HNE treatment increasedp53 and p21 expression and decreased expression of cyclinD2 The additional decrease of A- and B-cyclin suggests thatthe S- and G2-phase were also retarded contributing to theoverall slowdown of the cycle [326] In human breast cancercells (MCF7) the increase in endogenous levels of 4-HNEcaused by treatment with conjugated linoleic acid (CLA)resulted in the inhibition of cell proliferation through a p53-dependent mechanism [327] In human osteosarcoma cells(HOS) 4-HNE treatment declined gradually the proportionof cells in mitosis inhibited proliferation and differentiationand increased apoptosis [328] In malignant cells like hep-atome cells with a below-normal content of PUFAs and veryhigh expression of aldehyde dehydrogenase-3 (ADH3) whichmetabolize 4-HNE to DNH the inhibitory effects of 4-HNEon cell proliferation are lower but the inhibition of ADH3resulted in an increase in the quantity of aldehyde in the cellsand inhibit cell proliferation through the MAPK pathway byreduction of pRaf-1 and pERK12 [329 330] Moreover 4-HNE has also antiproliferativedifferentiative effect mainly inmalignant cell by affecting the expression of key genes suchas oncogenes (eg c-myc and c-myb) and cyclins In threehuman leukemic cell lines (HL-60 U937 andML-1) [313] andin colon cancer cells [265 314] cell proliferationwas inhibitedby 4-HNE as a consequence of downregulation of c-mycmRNA 4-HNE mediated inhibition of cell proliferation inthe HL-60 cell line by downregulation of Notch1 which isinvolved in expression of cyclin D1 and c-Myc [331] In SK-N-BE human neuroblastoma cells 4-HNE upregulated p53family gene expression and p53 gene targets p21 and bax andthe consequent reduction in S-phase cells and the increasedapoptotic cell proportion 4-HNE also reduced cyclin D2expression [332] In HepG2 cells 4-HNE decreased both cellsurvival and proliferation as evidenced by MTT assays andEdU incorporation as well as decreased expression of cyclinD1 and 120573-catenin [287] In K562 cells [333] HL-60 humanleukemic cell line [334] and murine erythroleukemia (MEL)cells [335] 4-HNE inhibited c-myc expression a oncogeneis involved in the regulation of cellular multiplication andtransformation (see review of Barrera and co-workers [336])


All these effects increased the proportion of G0G1 cellsindicating cell cycle arrest at G1 [324 325 336 337] 4-HNE-induced G2M cell cycle arrest was via p21 through amechanism (s) that is independent of p53The cell cycle arrestleads to apoptotic cell death [338] Enterococcus faecalismdashinfected macrophages produce 4-HNE This electrophilewhen purified mediated bystander effects in colonic epithe-lial cells by generating 120574H2AX foci and inducing G2M cellcycle arrest 4-HNE was also associated with mitotic spindledamage activation of stathmin cytokinesis failure and thedevelopment of tetraploid [339] In PC3 prostate cancer cell4-HNE induced G2M cell cycle arrest by decreasing p-Cdc2(entry into M phase is determined by activation of the Cdc2protein kinase which requires Cdc2 dephosphorylation)increased amount of p-H2AX indicated that 4-HNE inducedapoptotic cell death after a G2M accumulation [340]

In an opposite way different studies indicated that 4-HNE can promote cell proliferation in normal cells mainlyby upregulation of cyclin or E2F In cultured primary corticalneurons 4-HNE increased the protein levels of phospho-p53 and cell cycle-related proteins (cyclin D3 cyclin D1and CDC25A) caspase-3 activation PARP cleavage calpainactivation serinethreonine kinase 3 (Stk3) and sphingosinephosphate lyase 1 (Sgpl1) upregulation NAC decreased celldeath [341] In smooth muscle cells (SMCs) treatment with4-HNE enhanced cyclin D1 expression and activation of theERK signaling pathway which were stronger in young SMCscompared with aged SMCs [342] 4-HNE induced vascularsmooth muscle cell proliferation [142 343] Aldose reductase(AR) efficiently reduces 4-HNE and GS-HNE Inhibitionof AR can arrest cell cycle at S phase In VSMC cells theinhibition of AR prevents high glucose (HG-) andor TNF-alpha-induced VSMC proliferation by accumulating cells atthe G1 phase of the cell cycle Treatment of VSMC with 4-HNEor its glutathione conjugate (glutathionyl (GS-)HNE) orAR-catalyzed product of GS-HNE GS-14-dihydroxynonaneresulted in increased E2F-1 expression Inhibition of ARprevented 4-HNE- orGS-HNE-induced upregulation of E2F-1 Collectively these results show that AR could regulateHG- and TNF-alpha-inducedVSMCproliferation by alteringthe activation of G1S-phase proteins such as E2F-1 cdksand cyclins [344] In airway smooth muscle cells 4-HNEis mitogenic by increasing cyclin D1 activity through ERKsignaling pathway [345]

The differential effect of 4-HNE on cell proliferation inboth malignant and nonmalignant cells may be the conse-quence of lower aldehyde-metabolizing enzymes deregula-tion of antioxidant defenses and mitochondrial metabolismalteration [132 346] so that malignant cells are more vulner-able to further oxidative stress induced by exogenous ROS-generating agents or inhibitors of the antioxidant systems[347ndash349]

255 4-HNE-Induced Apoptosis and Necrosis Apoptosis isessential programmed cell death process for cells and itsdysregulation results in too little cell death which maycontribute to carcinogenesis or too much cell death whichmay be a component in the pathogenesis of several diseases

The alternative to apoptosis or programmed cell death isnecrosis or nonprogrammed cell death which is consideredto be a toxic process where the cell is a passive victim andfollows an energy-independent mode of death Dependingon the cell type DNA damagerepair capacity or cellularmetabolic circumstances 4-HNE can activate proliferativesignaling for cell division and promote cell survival or ldquostoprdquocell division and after prolonged arrest cells die fromapopto-sis 4-HNEmay induce these processes bymodulating severaltranscription factors sensible to stress such as Nrf2 AP-1 NF-120581B and PPAR or by modulating several signaling pathwaysincluding MAPK (p38 Erk and JNK) protein kinase Bprotein kinase C isoforms cell-cycle regulators receptortyrosine kinases and caspases Depending on 4-HNE con-centrations the cells ldquoendrdquo their lives by apoptosis or necrosisFor example the cytotoxicity of 4-HNE to HepG2 cells wasevaluated byMTTassay 4-HNEconcentrations ranging from10 to 100 120583M gradually decreased cell viability correspondingto an IC

50

value of 53 plusmn 239 120583M 4-HNE concentrationsof 5ndash40120583M caused apoptotic cell death (measured by flowcytometry caspase-3 activation and PARP cleavage) Finallya significant increase in necrotic cell population that is 318and 554 was observed in cells treated with 80 and 100 120583Mof 4-HNE respectively [350] These results show that 4-HNEinduces apoptosis at low concentration and necrosis at highconcentration

The two main pathways of apoptosis are extrinsic andintrinsic pathways The extrinsic signaling pathways thatinitiate apoptosis involve transmembrane receptor-mediatedinteractionsThis pathway is triggered by the binding of deathligands of the tumor necrosis factor (TNF) family to theirappropriate death receptors (DRs) on the cell surface best-characterized ligands and corresponding death receptorsinclude FasLFasR and TNF-120572TNFR1 [351 352] The intrin-sic signaling pathways that initiate apoptosis involve a diversearray of non-receptor-mediated stimuli The proapoptoticmember of the Bcl-2 family of proteins such as Bax per-meabilizes the outer mitochondrial membrane This allowsredistribution of cytochrome c from the mitochondrial inter-membrane space into the cytoplasm where it causes activa-tion of caspase proteases and subsequently cell death [352353] Each apoptosis pathway requires specific triggeringsignals to begin an energy-dependent cascade of molecularevents Each pathway activates its own initiator caspase (89) which in turn will activate the executioner caspase-3[352] The execution pathway results in characteristic cyto-morphological features including cell shrinkage chromatincondensation formation of cytoplasmic blebs and apoptoticbodies and finally phagocytosis of the apoptotic bodies byadjacent parenchymal cells neoplastic cells or macrophages[352 353] A multitude of mechanisms are employed byp53 to ensure efficient induction of apoptosis in a stage-tissue- and stress-signal-specific manner [354] 4-HNE-mediated activation of p53 may be one of the mechanismsresponsible for 4-HNE-induced apoptosis reported in manycell types For example in SH-SY5Y cells 4-HNE-inducedoxidative stress was associated with increased transcriptionaland translational expressions of Bax and p53 these eventstrigger other processes ending in cell death [355] In RPE


cells 4-HNE causes induction phosphorylation and nuclearaccumulation of p53 which is accompanied with downregu-lation of MDM2 a negative regulator of the p53 by blockingp53 transcriptional activity directly and mediating in thep53-degradation Associated proapoptotic genes Bax p21and JNK which are all signaling components p53-mediatedpathway of apoptosis are activated in response to exposure to4-HNE The induction of p53 by 4-HNE can be inhibited bythe overexpression of either hGSTA4 (in RPE cells) ormGsta4(in mice) which accelerates disposition of 4-HNE [356]In CRL25714 cell 4-HNE induced dose-dependent increasein the expression of p53 in the cytoplasmic and nuclearcompartments and increase in the expression of Bax [357]In human osteoarthritic chondrocytes 4-HNE treatment ledto p53 upregulation caspase-8 -9 and -3 activation Bcl-2 downregulation Bax upregulation cytochrome c-inducedrelease from mitochondria poly (ADP-ribose) polymerasecleavage DNA fragmentation FasCD95 upregulation Aktinhibition and energy depletion All these effects wereinhibited by an antioxidant N-acetyl-cysteine [358]

4-HNE can induce apoptosis through the death receptorFas (CD95-)mediated extrinsic pathway as well as throughthe p53-dependent intrinsic pathway For detailed infor-mation of the molecular mechanisms involved in 4-HNE-induced programmed cell death see review [359] Howeverthese mechanisms can be summarized in the following (i) 4-HNE is diffusible and can interact with Fas (CD95Apo1) onplasma membrane and upregulate and activate its expressionto mediate the apoptotic signaling through activation ofdownstream kinases (apoptosis signal-regulating kinase 1 orASK1 and JNK) which leads to activation of executionercaspase-3 and ending in apoptosis (ii) 4-HNE interacts withcytoplasmic p53 which causes its induction phosphoryla-tion and nuclear translocation In the nucleus p53 inhibitstranscription of antiapoptotic genes (Bcl2) and promotestranscription of proapoptotic genes (Bax) or cell cycle genes(p21) leading to activation of executioner caspase-3 andending in apoptosis or cell cycle arrest respectively (iii) 4-HNE also activates a negative feedback on Fas activationby a mechanism involving transcription repressor deathdomain-associated protein (Daxx) a nuclear protein which isassociated with DNA-binding transcription factors involvedin stress response 4-HNE interacts with the Daxx bound toheat shock factor-1 (HSF1) translocates Daxx from nucleusto cytoplasm where it binds to Fas and inhibits activation ofASK1 to limit apoptosis

256 4-HNE-Biomolecules Adducts The preference foramino acid modification by 4-HNE is Cys ≫ His gt Lysresulting in covalent adducts with the protein nucleophilicside chain [104 131 360 361] The reaction betweenprimary amines and 4-HNE carbonyl carbon groups yieldsa reversible Schiff base and the addition of thiol or aminocompounds on 4-HNE 120573-carbon atomrsquo (C of double bond)produces the corresponding Michael adduct [49] 4-HNE-protein adducts can contribute to protein crosslinking andinduce a carbonyl stress Recently it has been shown that amembrane associated protein called regulator of G-protein

signaling 4 (RGS4) can be modified by 4-HNE RGS4 likeother RGS proteins is responsible for temporally regulatingG-protein coupled receptor signaling by increasing theintrinsic GTPase activity of G120572 subunit of the heterotrimericsignaling complex 4-HNE modification of RGS4 at cysteineresidues during oxidative stress can disrupt RGS4 activityand alter signaling from stressed cells Possibly 4-HNE actsas an internal control for aberrant signaling due to excessRGS4 activity in a variety of pathologies where oxidativestress is a strong component [362] Our lab has reported that4-HNE can affect protein synthesis rates by forming adductwith eEF2 (see belowmdashcumene hydroperoxide-induced lipidperoxidation) Large lists of peptides and proteins known tobe modified by 4-HNE are given in the reviews [76 104 363]and including glutathione carnosine enzymatic proteinscarriers proteins membrane transport proteins receptorproteins cytoskeletal proteins chaperones mitochondrialupcoupling proteins transcription and protein synthesisfactors and antioxidant proteins

It has been reported that 4-HNE also could reactwith deoxyguanosine to form two pairs of diastereomeresadducts (4-HNE-dG 12 and 34) that further inducedDNA crosslink or DNA-protein conjugates The mecha-nism involves a nucleophilic Michael addition of the NH

2

-group of deoxyguanosine to the CC double bond of 4-HNE which yields 6-(1-hydroxyhexanyl)-8-hydroxy-1N(2)-propano-21015840-deoxyguanosine (HNE-dG) an exocyclic adduct[49 133 134] HNE-dG adducts have been detected in humanand animal tissues They are potentially mutagenic andcarcinogenic and can be repaired by the nucleotide excisionrepair (NER) pathway [364 365] In the presence of peroxidesa different reaction takes place and the stable end-productfound in the reaction of 4-HNE with DNA bases is etheno-DNA adducts because 4-HNE is converted by the peroxideto the corresponding epoxynonanal which then reacts tothe NH2-group of guanosine followed by cyclization reactionto form 1 N6-etheno-21015840-eoxyadenosine (120576dA) and 3 N4-etheno-21015840-deoxycytidine (120576dC) These 120576-adducts are elimi-nated by the base excision repair (BER) pathway [49 366]Etheno-DNA adduct levels were found to be significantlyelevated in the affected organs of subjects with chronicpancreatitis ulcerative colitis and Crohnrsquos disease whichprovide promising molecular signatures for risk predictionand potential targets and biomarkers for preventive measures[367 368] The 4-HNE-DNA adducts in tissue could serveas marker for the genetic damage produced by endogenousoxidation of omega-6-PUFAs

3 The Use of Mammalian Model in LipidPeroxidation Research CompoundsInduced Lipid Peroxidation

The use of mammalian model in lipid peroxidation researchis ideal for studying the consequences of lipid peroxidationin the context of whole organism and also to analyze theirinfluence on biomarkers to gain more insight into whatcontrols the lipid peroxidation and how lipid peroxidation-related diseases occur Animal models used to investigate the


+

LH

LH

Cumoxyl

Cumoperoxyl

Cumenehydroperoxide LP initiationpropagation

3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙

120572-Cumyl alcohol


Cumene hydroperoxide

L∙

L∙

O2

M(n+1)Mn

minusOH

Figure 7Mechanisms showing how cumene hydroperoxide produces lipophilic cumoxyl and cumoperoxyl radicals Cumene hydroperoxidein presence of transition metal ions produces cumoxyl radical (step 1) which abstracts a hydrogen (H) from a lipid (PUFA) molecule (LH)generating cumyl alcohol and lipid radical (L∙) that reacts readily with oxygen promoting the initiation or propagation of lipid peroxidation(Step 2) Cumoxyl radical can also react with other cumene hydroperoxide molecules to yield cumyl alcohol and cumoperoxyl radical (step3) Finally cumoperoxil radical may abstract hydrogen (H) from the closest available lipid to produce a new cumene hydroperoxide andlipid radical (L∙) which then again affects lipid peroxidation cycling (step 4) Cumoperoxyl radical may also react with oxygen to yield a newcumoxyl radical thus initiating a chain reaction (step 5)

genetic physiological or pathological consequences of lipidperoxidation should try to control the intrinsic and extrin-sic influences Genetic background diet environment andhealth status can be strictly controlled in many model organ-isms Compared with other model organisms such as worms(Caenorhabditis elegans) and flies (Drosophila melanogaster)the mammalian model is highly comparable to the humanin respect to organ systems tissues physiologic systems andeven behavioral traits Finally mammalian model in LP canbe used as a first step toward possible development of drugsor interventions to control lipid peroxidation process andprevent disease progression in humans Various mammalianmodels have been developed to study the lipid peroxidationprocess

31 Cumene Hydroperoxide-Induced Lipid PeroxidationCumene hydroperoxide (CH) a catalyst used in chemicaland pharmaceutical industry [369] is a stable organicoxidizing agent with the peroxy function group ndashOndashOndashwhich induces lipid peroxidation On the existence oftransition-metal CH can be reduced to form an alkoxylradical which can attack adjacent fatty acid side-chainsto produce lipid radical and cumyl alcohol The resultinglipid radical reacts with oxygen to form a lipid peroxylradical And a lipid peroxyl radical reacts with other fattyacid side-chains to produce a new lipid radical and lipidhydroperoxide and this chain reaction continues These lipidhydroperoxides may undergo transition-metal mediatedone-electron reduction and oxygenation to give lipid peroxylradicals which trigger exacerbating rounds of free radical-mediated lipid peroxidation (Figure 7) In our lab we have

made extensive use of membrane-soluble CH as a modelcompound for lipid hydroperoxides (LOOH) which areformed in the process of lipid peroxidation during oxidativestress CH-induced lipid peroxidation in animals has beenimportant to study the effect of lipid peroxidation on proteinsynthesis through mechanisms that involve regulation ofeElongation Factor 2 (eEF2) It is known that eEF2 plays akey role as a cytoplasmic component of the protein synthesismachinery where it is a fundamental regulatory protein ofthe translational elongation step that catalyzes the movementof the ribosome along the mRNA One particularity of eEF2is that it is quite sensitive to oxidative stress and is specificallyaffected by compounds that increase lipid peroxidationsuch as cumene hydroperoxide (CH) [370ndash373] We havepreviously reported that cytotoxic end-products of lipidperoxidation 4-HNE and MDA are able to form adductswith eEF2 in vitro [374] and in vivo [309] demonstrating forthe first time that this alteration of eEF2 could contributeto decline of protein synthesis secondary to LP increaseThe formation of these peroxide-eEF2-adducts is a possiblemechanism responsible of suboptimal hormone productionfrom hypothalamic-hypophysis system (HHS) duringoxidative stress and aging [375] The protection of eEF2alterations by end-products of lipid peroxidation must bespecifically carried out by compounds with lipoperoxylradical-scavenging features such as melatonin We havereported the ability of melatonin to protect against thechanges that occur in the eEF2 under conditions of lipidperoxidation induced by CH as well as decline of proteinsynthesis rate caused by lipid peroxidation demonstratingthat melatonin can prevent the decrease of several hormonesafter exposure to LP [376] In vitro studies carried out in


our lab also indicated that the antioxidants have differentcapacities to prevent eEF2 loss caused by CH [377 378] In rathippocampal neurons and in response to lipid peroxidationinduced by exposure to CH eEF2 subcellular localizationabundance and interaction with p53 were modified [379]Finally using CH-induced lipid peroxidation we foundthat a unique eEF2 posttranslational modified derivative ofhistidine (H715) known as diphthamide plays a role in theprotection of cells against the degradation of eEF2 and itis important to control the translation of IRES-dependentproteins XIAP and FGF2 two proteins that promote cellsurvival under conditions of oxidative stress [380] Otherlabs have used cumene hydroperoxide as a model compoundfor lipid hydroperoxides in vivo [381ndash385]

32 Tert Butyl Hydroperoxide It is an organic oxidizingagent containing a tertiary butyl group commonly used inindustry as prooxidizing a bleaching agent and an initiatorof polymerization Tert butyl hydroperoxide is a strongfree radical source and has been utilized to induce lipidperoxidation in vivomammalian model [386ndash392]

33 Carbon Tetrachloride (CCl4

) It is a toxic carcinogenicorganic compound which is used as a general solvent inindustrial degreasing operations It is also used as pesticidesand a chemical intermediate in the production of refrigerantsCarbon tetrachloride has been utilized to induce lipid perox-idation in vivomammalian model [90 393ndash398]

34 Quinolinic Acid (QA) It is a neuroactive metaboliteof the kynurenine pathway It is normally presented innanomolar concentrations in human brain and cerebrospinalfluid (CSF) and is often implicated in the pathogenesis ofa variety of human neurological diseases [399] QA hasbeen used to induce lipid peroxidation mediated by hydroxylradicals in vivomammalian models [400ndash405]

35 Transition Metals Ions They are essential elementswhich under certain conditions can have prooxidant effectRedox active transition metals have ability to induce andinitiate lipid peroxidation through the production of oxygenradicals mainly hydroxyl radical via FentonrsquosHaber-Weissreactions [63 406] Transition metal including copper [407ndash410] chromium [411 412] cadmium [413ndash416] nickel [417418] vanadium [419ndash421] manganese [59 422ndash424] andiron [59 407 425ndash434] has been utilized to induce lipidperoxidation in vivomammalian model

4 Pathological Processes Linked toMDA and 4-HNE

The accumulation of lipid peroxidation by-product has beenextensively studied and implicated in many toxic tissueinjuries and in pathological processes An increasing amountof literature has been published in the field In particular themeasurement of freeMDAandor 4-HNE levels or its derivedprotein adducts in biological samples from subjects affected

by several diseases has been widely utilized indirectly impli-catingMDAand 4-HNE in the pathogenesis of these diseasesTable 1 shows a brief extract of studies presented in theliterature in which MDA and 4-HNE have been found tobe significantly modified in pathological contexts The ldquobigrdquochallenge in the field of pathological processes is that it isoften difficult to determine whether these lipid peroxidation-derived aldehydes are actually involved in causing the diseaseor are a consequence to it

5 Conclusions

As conclusion in this review we summarized the physio-logical and pathophysiological role of lipid peroxides Whenoxidant compounds target lipids they can initiate the lipidperoxidation process a chain reaction that produces multiplebreakdown molecules such as MDA and 4-HNE Amongseveral substrates proteins and DNA are particularly suscep-tible to modification caused by these aldehydes MDA and 4-HNE adducts play a critical role inmultiple cellular processesand can participate in secondary deleterious reactions (egcrosslinking) by promoting intramolecular or intermolecularproteinDNA crosslinking that may induce profound alter-ation in the biochemical properties of biomolecules whichmay facilitate development of various pathological statesIdentification of specific aldehyde-modified molecules hasled to the determination of which selective cellular functionis altered For instance results obtained in our lab suggestthat lipid peroxidation affects protein synthesis in all tissuesduring aging through a mechanism involving the adductformation of MDA and 4-HNE with elongation factor-2However these molecules seem to have a dual behaviorsince cell response can tend to enhance survival or promotecell death depending of their cellular level and the pathwayactivated by them

Conflict of Interests

The authors declare no competing financial interests

Acknowledgments

This work was supported by Spanish Ministerio de Cienciae Innovacion BFU 2010 20882 and P10-CTS-6494 Mario FMunoz was supported by Consejeria de Economıa Innova-cion y Ciencia de la Junta de Andalucia (Spain) postdoctoralfellowship (P10-CTS-6494)

References

[1] G Fruhbeck J Gomez-Ambrosi F J Muruzabal and M ABurrell ldquoThe adipocyte a model for integration of endocrineand metabolic signaling in energy metabolism regulationrdquoTheAmerican Journal of Physiology Endocrinology andMetabolismvol 280 no 6 pp E827ndashE847 2001

[2] K N Frayn ldquoRegulation of fatty acid delivery in vivordquoAdvancesin ExperimentalMedicine andBiology vol 441 pp 171ndash179 1998

[3] E Vance and J E Vance Biochemistry Biochemistry of LipidsLipoproteins and Membranes 4th edition 2002


[4] K AMassey and A Nicolaou ldquoLipidomics of polyunsaturated-fatty-acid-derived oxygenatedmetabolitesrdquo Biochemical SocietyTransactions vol 39 no 5 pp 1240ndash1246 2011

[5] K AMassey and A Nicolaou ldquoLipidomics of oxidized polyun-saturated fatty acidsrdquo Free Radical Biology andMedicine vol 59pp 45ndash55 2013

[6] F R Jornayvaz and G I Shulman ldquoDiacylglycerol activationof protein kinase C120576 and hepatic insulin resistancerdquo CellMetabolism vol 15 no 5 pp 574ndash584 2012

[7] C Giorgi C Agnoletto C Baldini et al ldquoRedox control of pro-tein kinase C cell-and disease-specific aspectsrdquo Antioxidantsand Redox Signaling vol 13 no 7 pp 1051ndash1085 2010

[8] C Yang and M G Kazanietz ldquoChimaerins GAPs that bridgediacylglycerol signalling and the small G-protein Racrdquo Bio-chemical Journal vol 403 no 1 pp 1ndash12 2007

[9] J Baumann C Sevinsky and D S Conklin ldquoLipid biology ofbreast cancerrdquo Biochimica et Biophysica Acta vol 1831 no 10pp 1509ndash1517 2013

[10] S K Fisher J ENovak andBWAgranoff ldquoInositol and higherinositol phosphates in neural tissues homeostasis metabolismand functional significancerdquo Journal of Neurochemistry vol 82no 4 pp 736ndash754 2002

[11] S J Conway and G J Miller ldquoBiology-enabling inositolphosphates phosphatidylinositol phosphates and derivativesrdquoNatural Product Reports vol 24 no 4 pp 687ndash707 2007

[12] Y Takuwa Y Okamoto K Yoshioka and N TakuwaldquoSphingosine-1-phosphate signaling in physiology and dis-easesrdquo BioFactors vol 38 no 5 pp 329ndash337 2012

[13] M P Mattson Membrane Lipid Signaling in Aging and Age-Related Disease Elsevier 2003

[14] Y AHannun and LMObeid ldquoPrinciples of bioactive lipid sig-nalling lessons from sphingolipidsrdquo Nature Reviews MolecularCell Biology vol 9 no 2 pp 139ndash150 2008

[15] T Aoki and S Narumiya ldquoProstaglandins and chronic inflam-mationrdquo Trends in Pharmacological Sciences vol 33 no 6 pp304ndash311 2012

[16] E H C Tang P Libby P M Vanhoutte and A Xu ldquoAnti-inflammation therapy by activation of prostaglandin EP4 recep-tor in cardiovascular and other inflammatory diseasesrdquo Journalof Cardiovascular Pharmacology vol 59 no 2 pp 116ndash123 2012

[17] P Kalinski ldquoRegulation of immune responses by prostaglandinE2

rdquo Journal of Immunology vol 188 no 1 pp 21ndash28 2012[18] J G Kay and S Grinstein ldquoPhosphatidylserine-mediated cellu-

lar signalingrdquo Advances in Experimental Medicine and Biologyvol 991 pp 177ndash193 2013

[19] N Pluchino M Russo A N Santoro P Litta V Cela and AR Genazzani ldquoSteroid hormones and BDNFrdquoNeuroscience vol239 pp 271ndash279 2013

[20] L Moldovan and N I Moldovan ldquoOxygen free radicals andredox biology of organellesrdquo Histochemistry and Cell Biologyvol 122 no 4 pp 395ndash412 2004

[21] N Lane Oxygen The Molecule that Made the World OxfordUniversity Press 2002

[22] B Halliwell and J M C Gutteridge ldquoOxygen toxicity oxygenradicals transitionmetals anddiseaserdquoBiochemical Journal vol219 no 1 pp 1ndash14 1984

[23] J L VeneroM Revuelta L Atiki et al ldquoEvidence fordopamine-derived hydroxyl radical formation in the nigrostriatal systemin response to axotomyrdquo Free Radical Biology andMedicine vol34 no 1 pp 111ndash123 2003

[24] R J Castellani K Honda X Zhu et al ldquoContribution ofredox-active iron and copper to oxidative damage in Alzheimerdiseaserdquo Ageing Research Reviews vol 3 no 3 pp 319ndash3262004

[25] B Lipinski and E Pretorius ldquoHydroxyl radical-modified fib-rinogen as a marker of thrombosis the role of ironrdquo Hematol-ogy vol 17 no 4 pp 241ndash247 2012

[26] M Dizdaroglu and P Jaruga ldquoMechanisms of free radical-induced damage to DNArdquo Free Radical Research vol 46 no 4pp 382ndash419 2012

[27] T Kanno K Nakamura H Ikai K Kikuchi K Sasaki and YNiwano ldquoLiterature review of the role of hydroxyl radicals inchemically-induced mutagenicity and carcinogenicity for therisk assessment of a disinfection system utilizing photolysisof hydrogen peroxiderdquo Journal of Clinical Biochemistry andNutrition vol 51 no 1 pp 9ndash14 2012

[28] B H J Bielski R L Arudi and M W Sutherland ldquoA study ofthe reactivity of HO2O2- with unsaturated fatty acidsrdquo Journalof Biological Chemistry vol 258 no 8 pp 4759ndash4761 1983

[29] C Schneider W E Boeglin H Yin N A Porter and A RBrash ldquoIntermolecular peroxyl radical reactions during autoxi-dation of hydroxy and hydroperoxy arachidonic acids generatea novel series of epoxidized productsrdquo Chemical Research inToxicology vol 21 no 4 pp 895ndash903 2008

[30] R W Browne and D Armstrong ldquoHPLC analysis of lipid-derived polyunsaturated fatty acid peroxidation products inoxidatively modified human plasmardquo Clinical Chemistry vol46 no 6 part 1 pp 829ndash836 2000

[31] H Yin L Xu and N A Porter ldquoFree radical lipid peroxidationmechanisms and analysisrdquoChemical Reviews vol 111 no 10 pp5944ndash5972 2011

[32] R Volinsky and P K J Kinnunen ldquoOxidized phosphatidyl-cholines in membrane-level cellular signaling from biophysicsto physiology andmolecular pathologyrdquo FEBS Journal vol 280no 12 pp 2806ndash2816 2013

[33] P K J Kinnunen K Kaarniranta and A K Mahalka ldquoProtein-oxidized phospholipid interactions in cellular signaling for celldeath from biophysics to clinical correlationsrdquo Biochimica etBiophysica Acta vol 1818 no 10 pp 2446ndash2455 2012

[34] A Reis and C M Spickett ldquoChemistry of phospholipid oxida-tionrdquo Biochimica et Biophysica Acta vol 1818 no 10 pp 2374ndash2387 2012

[35] G O Fruhwirth A Loidl and A Hermetter ldquoOxidized phos-pholipids from molecular properties to diseaserdquo Biochimica etBiophysica Acta Molecular Basis of Disease vol 1772 no 7 pp718ndash736 2007

[36] A W Girotti ldquoLipid hydroperoxide generation turnover andeffector action in biological systemsrdquo Journal of Lipid Researchvol 39 no 8 pp 1529ndash1542 1998

[37] J Kanner J B German and J E Kinsella ldquoInitiation of lipidperoxidation in biological systemsrdquo Critical Reviews in FoodScience and Nutrition vol 25 no 4 pp 317ndash364 1987

[38] H Esterbauer K H Cheeseman and M U Dianzani ldquoSepa-ration and characterization of the aldehydic products of lipidperoxidation stimulated by ADP-Fe2+ in rat liver microsomesrdquoBiochemical Journal vol 208 no 1 pp 129ndash140 1982

[39] G Poli M U Dianzani K H Cheeseman T F Slater J Langand H Esterbauer ldquoSeparation and characterization of thealdehydic products of lipid peroxidation stimulated by carbontetrachloride or ADP-iron in isolated rat hepatocytes and ratlivermicrosomal suspensionsrdquoBiochemical Journal vol 227 no2 pp 629ndash638 1985


[40] A Benedetti M Comporti and H Esterbauer ldquoIdentificationof 4-hydroxynonenal as a cytotoxic product originating fromthe peroxidation of liver microsomal lipidsrdquo Biochimica etBiophysica Acta vol 620 no 2 pp 281ndash296 1980

[41] E Cadenas A Muller R Brigelius H Esterbauer and H SiesldquoEffects of 4-hydroxynonenal on isolated hepatocytes Studieson chemiluminescence response alkane production and glu-tathione statusrdquoBiochemical Journal vol 214 no 2 pp 479ndash4871983

[42] H Esterbauer J Lang S Zadravec andT F Slater ldquoDetection ofmalonaldehyde by high-performance liquid chromatographyrdquoMethods in Enzymology vol 105 pp 319ndash328 1984

[43] P Winkler W Lindner H Esterbauer E Schauenstein R JSchaur and G A Khoschsorur ldquoDetection of 4-hydro-xynonenal as a product of lipid peroxidation in native Ehrlichascites tumor cellsrdquo Biochimica et Biophysica Acta Lipids andLipid Metabolism vol 796 no 3 pp 232ndash237 1984

[44] H Esterbauer A Benedetti J Lang R Fulceri G Faulerand M Comporti ldquoStudies on the mechanism of formationof 4-hydroxynonenal during microsomal lipid peroxidationrdquoBiochimica et Biophysica Acta Lipids and Lipid Metabolism vol876 no 1 pp 154ndash166 1986

[45] J S Hurst T F Slater and J Lang ldquoEffects of the lipidperoxidation product 4-hydroxynonenal on the aggregation ofhuman plateletsrdquo Chemico-Biological Interactions vol 61 no 2pp 109ndash124 1987

[46] K H Cheeseman A Beavis and H Esterbauer ldquoHydroxyl-radical-induced iron-catalysed degradation of 2-deoxyriboseQuantitative determination of malondialdehyderdquo BiochemicalJournal vol 252 no 3 pp 649ndash653 1988

[47] H Esterbauer and H Zolliner ldquoMethods for determination ofaldehydic lipid peroxidation productsrdquo Free Radical Biology andMedicine vol 7 no 2 pp 197ndash203 1989

[48] H Esterbauer and K H Cheeseman ldquoDetermination of alde-hydic lipid peroxidation products malonaldehyde and 4-hydroxynonenalrdquoMethods in Enzymology vol 186 pp 407ndash4211990

[49] H Esterbauer R J Schaur and H Zollner ldquoChemistry andBiochemistry of 4-hydroxynonenal malonaldehyde and relatedaldehydesrdquo Free Radical Biology and Medicine vol 11 no 1 pp81ndash128 1991

[50] H Esterbauer P Eckl and A Ortner ldquoPossible mutagensderived from lipids and lipid precursorsrdquo Mutation Researchvol 238 no 3 pp 223ndash233 1990

[51] W A Pryor ldquoOn the detection of lipid hydroperoxides inbiological samplesrdquo Free Radical Biology and Medicine vol 7no 2 pp 177ndash178 1989

[52] R O Sinnhuber T C Yu and T C Yu ldquoCharacterization of thered pigment formed in the 2-thiobarbituric acid determinationof oxidative rancidityrdquo Journal of Food Science vol 23 no 6 pp626ndash634 1958

[53] MGieraH Lingeman andWMANiessen ldquoRecent advance-ments in the LC- and GC-based analysis of malondialdehyde(MDA) a brief overviewrdquo Chromatographia vol 75 no 9-10pp 433ndash440 2012

[54] E Schauenstein ldquoAutoxidation of polyunsaturated esters inwater chemical structure and biological activity of the prod-uctsrdquo Journal of Lipid Research vol 8 no 5 pp 417ndash428 1967

[55] G Brambilla L Sciaba P Faggin et al ldquoCytotoxicity DNAfragmentation and sister-chromatid exchange in Chinese ham-ster ovary cells exposed to the lipid peroxidation prod-uct 4-hydroxynonenal and homologous aldehydesrdquo MutationResearch vol 171 no 2-3 pp 169ndash176 1986

[56] R J Schaur ldquoBasic aspects of the biochemical reactivity of 4-hydroxynonenalrdquoMolecular Aspects of Medicine vol 24 no 4-5 pp 149ndash159 2003

[57] N Zarkovic ldquo4-Hydroxynonenal as a bioactive marker ofpathophysiological processesrdquo Molecular Aspects of Medicinevol 24 no 4-5 pp 281ndash291 2003

[58] E Niki ldquoBiomarkers of lipid peroxidation in clinical materialrdquoBiochimica et Biophysica Acta vol 1840 no 2 pp 809ndash817 2014

[59] S Arguelles S Garcıa M Maldonado A Machado and AAyala ldquoDo the serum oxidative stress biomarkers providea reasonable index of the general oxidative stress statusrdquoBiochimica et Biophysica Acta General Subjects vol 1674 no3 pp 251ndash259 2004

[60] S Arguelles A Gomez A Machado and A Ayala ldquoA pre-liminary analysis of within-subject variation in human serumoxidative stress parameters as a function of timerdquo RejuvenationResearch vol 10 no 4 pp 621ndash636 2007

[61] R Brigelius-Flohe andMMaiorino ldquoGlutathione peroxidasesrdquoBiochimica et Biophysica Acta vol 1830 no 5 pp 3289ndash33032013

[62] H Steinbrenner and H Sies ldquoProtection against reactive oxy-gen species by selenoproteinsrdquo Biochimica et Biophysica ActaGeneral Subjects vol 1790 no 11 pp 1478ndash1485 2009

[63] M Valko H Morris andM T D Cronin ldquoMetals toxicity andoxidative stressrdquoCurrentMedicinal Chemistry vol 12 no 10 pp1161ndash1208 2005

[64] C Szabo H Ischiropoulos and R Radi ldquoPeroxynitrite bio-chemistry pathophysiology and development of therapeuticsrdquoNature Reviews Drug Discovery vol 6 no 8 pp 662ndash680 2007

[65] C CWinterbourn ldquoBiological reactivity and biomarkers of theneutrophil oxidant hypochlorous acidrdquo Toxicology vol 181-182pp 223ndash227 2002

[66] E Malle G Marsche J Arnhold and M J Davies ldquoModifi-cation of low-density lipoprotein by myeloperoxidase-derivedoxidants and reagent hypochlorous acidrdquoBiochimica et Biophys-ica ActaMolecular and Cell Biology of Lipids vol 1761 no 4 pp392ndash415 2006

[67] S Miyamoto G E Ronsein F M Prado et al ldquoBiologi-cal hydroperoxides and singlet molecular oxygen generationrdquoIUBMB Life vol 59 no 4-5 pp 322ndash331 2007

[68] S Miyamoto G R Martinez D Rettori O Augusto M H GMedeiros and PDiMascio ldquoLinoleic acid hydroperoxide reactswith hypochlorous acid generating peroxyl radical intermedi-ates and singlet molecular oxygenrdquo Proceedings of the NationalAcademy of Sciences of the United States of America vol 103 no2 pp 293ndash298 2006

[69] M Gracanin C L Hawkins D I Pattison and M J DaviesldquoSinglet-oxygen-mediated amino acid and protein oxidationformation of tryptophan peroxides and decomposition prod-uctsrdquo Free Radical Biology and Medicine vol 47 no 1 pp 92ndash102 2009

[70] M J Davies ldquoSinglet oxygen-mediated damage to proteinsand its consequencesrdquo Biochemical and Biophysical ResearchCommunications vol 305 no 3 pp 761ndash770 2003

[71] R M Domingues P Domingues T Melo D Perez-Sala AReis and C M Spickett ldquoLipoxidation adducts with peptides


and proteins deleterious modifications or signaling mecha-nismsrdquo Journal of Proteomics vol 92 pp 110ndash131 2013

[72] A Negre-Salvayre C Coatrieux C Ingueneau and R SalvayreldquoAdvanced lipid peroxidation end products in oxidative damageto proteins Potential role in diseases and therapeutic prospectsfor the inhibitorsrdquo British Journal of Pharmacology vol 153 no1 pp 6ndash20 2008

[73] X Wang X G Lei and J Wang ldquoMalondialdehyde regu-lates glucose-stimulated insulin secretion in murine islets viaTCF7L2-dependent Wnt signaling pathwayrdquo Molecular andCellular Endocrinology vol 382 no 1 pp 8ndash16 2014

[74] I Garcıa-Ruiz P de la Torre T Dıaz et al ldquoSp1 and Sp3 tran-scription factors mediate malondialdehyde-induced collagenalpha 1(I) gene expression in cultured hepatic stellate cellsrdquoTheJournal of Biological Chemistry vol 277 no 34 pp 30551ndash305582002

[75] L Li and J R Davie ldquoThe role of Sp1 and Sp3 in normal andcancer cell biologyrdquoAnnals of Anatomy Anatomischer Anzeigervol 192 no 5 pp 275ndash283 2010

[76] N Zarkovic A Cipak M Jaganjac S Borovic and K ZarkovicldquoPathophysiological relevance of aldehydic protein modifica-tionsrdquo Journal of Proteomics vol 92 pp 239ndash247 2013

[77] I A Blair ldquoDNA adducts with lipid peroxidation productsrdquoJournal of Biological Chemistry vol 283 no 23 pp 15545ndash155492008

[78] W Łuczaj and E Skrzydlewska ldquoDNA damage caused by lipidperoxidation productsrdquo Cellular and Molecular Biology Lettersvol 8 no 2 pp 391ndash413 2003

[79] S C Garcia D Grotto R P Bulcao et al ldquoEvaluation of lipiddamage related to pathological and physiological conditionsrdquoDrug and Chemical Toxicology vol 36 no 3 pp 306ndash312 2013

[80] G Li Y Chen H Hu et al ldquoAssociation between age-relateddecline of kidney function and plasma malondialdehyderdquoRejuvenation Research vol 15 no 3 pp 257ndash264 2012

[81] J Sanyal S K Bandyopadhyay T K Banerjee et al ldquoPlasmalevels of lipid peroxides in patients with Parkinsonrsquos diseaserdquoEuropean Review for Medical and Pharmacological Sciences vol13 no 2 pp 129ndash132 2009

[82] N Shanmugam J L Figarola Y Li P M Swiderski S Rahbarand R Natarajan ldquoProinflammatory effects of advanced lipoxi-dation end products in monocytesrdquo Diabetes vol 57 no 4 pp879ndash888 2008

[83] G Baskol H Demir M Baskol et al ldquoInvestigation of proteinoxidation and lipid peroxidation in patients with rheumatoidarthritisrdquoCell Biochemistry and Function vol 24 no 4 pp 307ndash311 2006

[84] R A Merendino F Salvo A Saija et al ldquoMalondialdehyde inbenign prostate hypertrophy a useful markerrdquo Mediators ofInflammation vol 12 no 2 pp 127ndash128 2003

[85] P L Paggiaro M L Bartoli F Novelli et al ldquoMalondialdehydein exhaled breath condensate as a marker of oxidative stress indifferent pulmonary diseasesrdquo Mediators of Inflammation vol2011 Article ID 891752 7 pages 2011

[86] M Hecker and V Ullrich ldquoOn the mechanism of prostacyclinand thromboxane A2 biosynthesisrdquo Journal of Biological Chem-istry vol 264 no 1 pp 141ndash150 1989

[87] R A Sharma A Gescher J P Plastaras et al ldquoCyclooxygenase-2 malondialdehyde and pyrimidopurinone adducts ofdeoxyguanosine in human colon cellsrdquo Carcinogenesis vol 22no 9 pp 1557ndash1560 2001

[88] D Tsikas M T Suchy J Niemann et al ldquoGlutathione promotesprostaglandin H synthase (cyclooxygenase)-dependent forma-tion of malondialdehyde and 15(S)-8-iso-prostaglandin F2120572rdquoFEBS Letters vol 586 no 20 pp 3723ndash3730 2012

[89] M Griesser W E Boeglin T Suzuki and C SchneiderldquoConvergence of the 5-LOX and COX-2 pathways heme-catalyzed cleavage of the 5S-HETE-derived di-endoperoxideinto aldehyde fragmentsrdquo Journal of Lipid Research vol 50 no12 pp 2455ndash2462 2009

[90] M B Kadiiska B C Gladen D D Baird et al ldquoBiomarkersof oxidative stress study III Effects of the nonsteroidal anti-inflammatory agents indomethacin and meclofenamic acidon measurements of oxidative products of lipids in CCl4poisoningrdquo Free Radical Biology andMedicine vol 38 no 6 pp711ndash718 2005

[91] E Ricciotti and G A FitzGerald ldquoProstaglandins and inflam-mationrdquo Arteriosclerosis Thrombosis and Vascular Biology vol31 no 5 pp 986ndash1000 2011

[92] P Ekambaram W Lambiv R Cazzolli A W Ashton and KV Honn ldquoThe thromboxane synthase and receptor signalingpathway in cancer an emerging paradigm in cancer progressionandmetastasisrdquo Cancer andMetastasis Reviews vol 30 no 3-4pp 397ndash408 2011

[93] H Yang andC Chen ldquoCyclooxygenase-2 in synaptic signalingrdquoCurrent Pharmaceutical Design vol 14 no 14 pp 1443ndash14512008

[94] W A Pryor J P Stanley and E Blair ldquoAutoxidation ofpolyunsaturated fatty acids II A suggested mechanism for theformation of TBA reactive materials from prostaglandin likeendoperoxidesrdquo Lipids vol 11 no 5 pp 370ndash379 1976

[95] G L Milne H Yin and J D Morrow ldquoHuman biochemistry ofthe isoprostane pathwayrdquo Journal of Biological Chemistry vol283 no 23 pp 15533ndash15537 2008

[96] L Gao W E Zackert J J Hasford et al ldquoFormation ofprostaglandins E2 and D2 via the isoprostane pathway Amechanism for the generation of bioactive prostaglandins inde-pendent of cyclooxygenaserdquo Journal of Biological Chemistry vol278 no 31 pp 28479ndash28489 2003

[97] H Yin L Gao H-H Tai L J Murphey N A Porter and JD Morrow ldquoUrinary prostaglandin F2120572 is generated from theisoprostane pathway and not the cyclooxygenase in humansrdquoJournal of Biological Chemistry vol 282 no 1 pp 329ndash336 2007

[98] J D Brooks G L Milne H Yin S C Sanchez N A Porterand J DMorrow ldquoFormation of highly reactive cyclopentenoneisoprostane compounds (A 3J3-isoprostanes) in vivo fromeicosapentaenoic acidrdquo Journal of Biological Chemistry vol 283no 18 pp 12043ndash12055 2008

[99] L J Roberts II J P Fessel and S S Davies ldquoThe biochemistry ofthe isoprostane neuroprostane and isofuran pathways of lipidperoxidationrdquo Brain Pathology vol 15 no 2 pp 143ndash148 2005

[100] A N Onyango and N Baba ldquoNew hypotheses on the pathwaysof formation of malondialdehyde and isofuransrdquo Free RadicalBiology and Medicine vol 49 no 10 pp 1594ndash1600 2010

[101] G M Siu and H H Draper ldquoMetabolism of malonaldehyde invivo and in vitrordquo Lipids vol 17 no 5 pp 349ndash355 1982

[102] L J Marnett J Buck M A Tuttle A K Basu and A WBull ldquoDistribution and oxidation of malondialdehyde in micerdquoProstaglandins vol 30 no 2 pp 241ndash254 1985

[103] Z S Agadjanyan L F Dmitriev and S F Dugin ldquoA new roleof phosphoglucose isomerase Involvement of the glycolyticenzyme in aldehyde metabolismrdquo Biochemistry vol 70 no 11pp 1251ndash1255 2005


[104] S Pizzimenti E Ciamporcero M Daga et al ldquoInteractionof aldehydes derived from lipid peroxidation and membraneproteinsrdquo Frontiers in Physiology vol 4 article 242 2013

[105] A Skoumalova and J Hort ldquoBlood markers of oxidative stressin Alzheimerrsquos diseaserdquo Journal of Cellular and MolecularMedicine vol 16 no 10 pp 2291ndash2300 2012

[106] F Mangialasche M C Polidori R Monastero et al ldquoBiomark-ers of oxidative and nitrosative damage in Alzheimerrsquos diseaseand mild cognitive impairmentrdquo Ageing Research Reviews vol8 no 4 pp 285ndash305 2009

[107] R Pamplona E Dalfo V Ayala et al ldquoProteins in humanbrain cortex are modified by oxidation glycoxidation andlipoxidation effects of Alzheimer disease and identification oflipoxidation targetsrdquo Journal of Biological Chemistry vol 280no 22 pp 21522ndash21530 2005

[108] D O Cristalli N Arnal F A Marra M J T De Alaniz andC AMarra ldquoPeripheral markers in neurodegenerative patientsand their first-degree relativesrdquo Journal of the NeurologicalSciences vol 314 no 1-2 pp 48ndash56 2012

[109] M Valko D Leibfritz J Moncol M T D Cronin M Mazurand J Telser ldquoFree radicals and antioxidants in normal physi-ological functions and human diseaserdquo International Journal ofBiochemistry and Cell Biology vol 39 no 1 pp 44ndash84 2007

[110] N Lopez C Tormo I De Blas I Llinares and J AlomldquoOxidative stress in Alzheimerrsquos disease and mild cognitiveimpairment with high sensitivity and specificityrdquo Journal ofAlzheimerrsquos Disease vol 33 no 3 pp 823ndash829 2013

[111] L L Torres N B Quaglio G T De Souza et al ldquoPeripheraloxidative stress biomarkers in mild cognitive impairment andalzheimerrsquos diseaserdquo Journal of Alzheimerrsquos Disease vol 26 no1 pp 59ndash68 2011

[112] M C Polidori and P Mecocci ldquoPlasma susceptibility to freeradical-induced antioxidant consumption and lipid peroxida-tion is increased in very old subjects with Alzheimer diseaserdquoJournal of Alzheimerrsquos Disease vol 4 no 6 pp 517ndash522 2002

[113] M Padurariu A Ciobica L Hritcu B Stoica W Bild andC Stefanescu ldquoChanges of some oxidative stress markers inthe serum of patients with mild cognitive impairment andAlzheimerrsquos diseaserdquoNeuroscience Letters vol 469 no 1 pp 6ndash10 2010

[114] L H Sanders and J Timothy Greenamyre ldquoOxidative dam-age to macromolecules in human Parkinson disease and therotenone modelrdquo Free Radical Biology andMedicine vol 62 pp111ndash120 2013

[115] R B Mythri C Venkateshappa G Harish et al ldquoEvaluation ofMarkers of oxidative stress antioxidant function and astrocyticproliferation in the striatum and frontal cortex of Parkinsonrsquosdisease brainsrdquoNeurochemical Research vol 36 no 8 pp 1452ndash1463 2011

[116] A Navarro A Boveris M J Bandez et al ldquoHuman braincortex mitochondrial oxidative damage and adaptive responsein Parkinson disease and in dementia with Lewy bodiesrdquo FreeRadical Biology and Medicine vol 46 no 12 pp 1574ndash15802009

[117] A Kilinc A S Yalcin D Yalcin Y Taga and K EmerkldquoIncreased erythrocyte susceptibility to lipid peroxidation inhuman Parkinsonrsquos diseaserdquo Neuroscience Letters vol 87 no 3pp 307ndash310 1988

[118] A Baillet V Chanteperdrix C Trocme P Casez C Garrel andG Besson ldquoThe role of oxidative stress in amyotrophic lateralsclerosis and Parkinsonrsquos diseaserdquo Neurochemical Research vol35 no 10 pp 1530ndash1537 2010

[119] C M Chen J L Liu Y R Wu et al ldquoIncreased oxidative dam-age in peripheral blood correlates with severity of Parkinsonrsquosdiseaserdquo Neurobiology of Disease vol 33 no 3 pp 429ndash4352009

[120] J Kalra A H Rajput S V Mantha A K Chaudhary andK Prasad ldquoOxygen free radical producing activity of poly-morphonuclear leukocytes in patients with Parkinsonrsquos diseaserdquoMolecular and Cellular Biochemistry vol 112 no 2 pp 181ndash1861992

[121] S Younes-Mhenni M Frih-Ayed A Kerkeni M Bost andG Chazot ldquoPeripheral blood markers of oxidative stress inParkinsonrsquos diseaserdquo European Neurology vol 58 no 2 pp 78ndash83 2007

[122] L J Niedernhofer J S Daniels C A Rouzer R E Greeneand L J Marnett ldquoMalondialdehyde a product of lipid per-oxidation is mutagenic in human cellsrdquo Journal of BiologicalChemistry vol 278 no 33 pp 31426ndash31433 2003

[123] D Del Rio A J Stewart and N Pellegrini ldquoA review ofrecent studies on malondialdehyde as toxic molecule andbiologicalmarker of oxidative stressrdquoNutritionMetabolism andCardiovascular Diseases vol 15 no 4 pp 316ndash328 2005

[124] L A VanderVeen M F Hashim Y Shyr and L J MarnettldquoInduction of frameshift and base pair substitution mutationsby the major DNA adduct of the endogenous carcinogen mal-ondialdehyderdquo Proceedings of the National Academy of Sciencesof the United States of America vol 100 no 24 pp 14247ndash142522003

[125] M E M Peluso A Munnia V Bollati et al ldquoAberrant methy-lation of hypermethylated-in-cancer-1 and exocyclic DNAadducts in tobacco smokersrdquo Toxicological Sciences vol 137 no1 pp 47ndash54 2014

[126] F Cai Y M Dupertuis and C Pichard ldquoRole of polyunsat-urated fatty acids and lipid peroxidation on colorectal cancerrisk and treatmentsrdquo Current Opinion in Clinical Nutrition andMetabolic Care vol 15 no 2 pp 99ndash106 2012

[127] U Nair H Bartsch and J Nair ldquoLipid peroxidation-inducedDNA damage in cancer-prone inflammatory diseases a reviewof published adduct types and levels in humansrdquo Free RadicalBiology and Medicine vol 43 no 8 pp 1109ndash1120 2007

[128] H Bartsch and J Nair ldquoAccumulation of lipid peroxidation-derived DNA lesions potential lead markers for chemopreven-tion of inflammation-driven malignanciesrdquoMutation ResearchFundamental and Molecular Mechanisms of Mutagenesis vol591 no 1-2 pp 34ndash44 2005

[129] M Wang K Dhingra W N Hittelman J G Liehr M DeAndrade and D Li ldquoLipid peroxidation-induced putativemalondialdehyde-DNA adducts in human breast tissuesrdquo Can-cer Epidemiology Biomarkers and Prevention vol 5 no 9 pp705ndash710 1996

[130] L J Marnett ldquoInflammation and cancer chemical approachesto mechanisms imaging and treatmentrdquo Journal of OrganicChemistry vol 77 no 12 pp 5224ndash5238 2012

[131] S Dalleau M Baradat F Gueraud and L Huc ldquoCell death anddiseases related to oxidative stress 4-hydroxynonenal (HNE) inthe balancerdquo Cell Death and Differentiation vol 20 no 12 pp1615ndash1630 2013

[132] G Barrera ldquoOxidative stress and lipid peroxidation productsin cancer progression and therapyrdquo ISRN Oncology vol 2012Article ID 137289 21 pages 2012

[133] H Huang I D Kozekov A Kozekova et al ldquoDNA cross-link induced by trans-4-hydroxynonenalrdquo Environmental andMolecular Mutagenesis vol 51 no 6 pp 625ndash634 2010


[134] I G Minko I D Kozekov T M Harris C J Rizzo R S Lloydand M P Stone ldquoChemistry and biology of DNA containing1N2-deoxyguanosine adducts of the120572120573-unsaturated aldehydesacrolein crotonaldehyde and 4-hydroxynonenalrdquo ChemicalResearch in Toxicology vol 22 no 5 pp 759ndash778 2009

[135] ANegre-Salvayre N Auge V Ayala et al ldquoPathological aspectsof lipid peroxidationrdquo Free Radical Research vol 44 no 10 pp1125ndash1171 2010

[136] F-L Chung J Pan S Choudhury R Roy W Hu and M-S Tang ldquoFormation of trans-4-hydroxy-2-nonenal- and otherenal-derived cyclic DNA adducts from 120596-3 and 120596-6 polyunsat-urated fatty acids and their roles in DNA repair and human p53genemutationrdquoMutationResearch Fundamental andMolecularMechanisms of Mutagenesis vol 531 no 1-2 pp 25ndash36 2003

[137] R Lee M Margaritis K M Channon and C AntoniadesldquoEvaluating oxidative stress in human cardiovascular diseasemethodological aspects and considerationsrdquo Current MedicinalChemistry vol 19 no 16 pp 2504ndash2520 2012

[138] K Uchida ldquoRole of reactive aldehyde in cardiovascular dis-easesrdquo Free Radical Biology and Medicine vol 28 no 12 pp1685ndash1696 2000

[139] E J Anderson L A Katunga and M S Willis ldquoMitochondriaas a source and target of lipid peroxidation products in healthyand diseased heartrdquo Clinical and Experimental Pharmacologyand Physiology vol 39 no 2 pp 179ndash193 2012

[140] E U Nwose H F Jelinek R S Richards and R G Kerr ldquoEry-throcyte oxidative stress in clinical management of diabetes andits cardiovascular complicationsrdquo British Journal of BiomedicalScience vol 64 no 1 pp 35ndash43 2007

[141] E Ho K Karimi Galougahi C C Liu R Bhindi and G AFigtree ldquoBiological markers of oxidative stress applications tocardiovascular research and practicerdquo Redox Biology vol 1 no1 pp 483ndash491 2013

[142] S J Chapple X Cheng and G E Mann ldquoEffects of 4-hydroxynonenal on vascular endothelial and smooth musclecell redox signaling and function in health and diseaserdquo RedoxBiology vol 1 no 1 pp 319ndash331 2013

[143] M P Mattson ldquoRoles of the lipid peroxidation product 4-hydroxynonenal in obesity the metabolic syndrome and asso-ciated vascular and neurodegenerative disordersrdquo ExperimentalGerontology vol 44 no 10 pp 625ndash633 2009

[144] G Leonarduzzi E Chiarpotto F Biasi and G Poli ldquo4-Hydroxynonenal and cholesterol oxidation products inatherosclerosisrdquo Molecular Nutrition and Food Research vol49 no 11 pp 1044ndash1049 2005

[145] D A Slatter C H Bolton and A J Bailey ldquoThe importance oflipid-derived malondialdehyde in diabetes mellitusrdquoDiabetolo-gia vol 43 no 5 pp 550ndash557 2000

[146] Y Bhutia A Ghosh M L Sherpa R Pal and P K MohantaldquoSerum malondialdehyde level surrogate stress marker in theSikkimese diabeticsrdquo Journal of Natural Science Biology andMedicine vol 2 no 1 pp 107ndash112 2011

[147] R Mahreen M Mohsin Z Nasreen M Siraj and M IshaqldquoSignificantly increased levels of serum malonaldehyde in type2 diabetics with myocardial infarctionrdquo International Journal ofDiabetes in Developing Countries vol 30 no 1 pp 49ndash51 2010

[148] B K Tiwari K B Pandey A B Abidi and S I RizvildquoMarkers of oxidative stress during diabetes mellitusrdquo Journalof Biomarkers vol 2013 Article ID 378790 8 pages 2013

[149] M Nakhjavani A Esteghamati S Nowroozi F Asgarani ARashidi andOKhalilzadeh ldquoType 2 diabetesmellitus duration

an independent predictor of serum malondialdehyde levelsrdquoSingapore Medical Journal vol 51 no 7 pp 582ndash585 2010

[150] C H Wang R W Chang Y H Ko et al ldquoPrevention ofarterial stiffening by using low-dose atorvastatin in diabetes isassociated with decreased malondialdehyderdquo PloS ONE vol 9no 3 Article ID e90471 2014

[151] M Jaganjac O Tirosh G Cohen S Sasson and N ZarkovicldquoReactive aldehydesmdashsecond messengers of free radicals indiabetes mellitusrdquo Free Radical Research vol 47 supplement 1pp 39ndash48 2013

[152] G Cohen Y Riahi O Shamni et al ldquoRole of lipid peroxi-dation and PPAR-120575 in amplifying glucose-stimulated insulinsecretionrdquo Diabetes vol 60 no 11 pp 2830ndash2842 2011

[153] A R Pradeep E Agarwal P Bajaj and N S Rao ldquo4-Hydroxy-2-nonenal an oxidative stress marker in crevicularfluid and serum in type 2 diabetes with chronic periodontitisrdquoContemporary Clinical Dentistry vol 4 no 3 pp 281ndash285 2013

[154] S Toyokuni S Yamada M Kashima et al ldquoSerum 4-hydroxy-2-nonenal-modified albumin is elevated in patients with type 2diabetes mellitusrdquo Antioxidants and Redox Signaling vol 2 no4 pp 681ndash685 2000

[155] G Cohen Y Riahi V Sunda et al ldquoSignaling properties of 4-hydroxyalkenals formed by lipid peroxidation in diabetesrdquo FreeRadical Biology and Medicine vol 65 pp 978ndash987 2013

[156] S Lupachyk H Shevalye Y Maksimchyk V R Drel andI G Obrosova ldquoPARP inhibition alleviates diabetes-inducedsystemic oxidative stress and neural tissue 4-hydroxynonenaladduct accumulation correlation with peripheral nerve func-tionrdquo Free Radical Biology and Medicine vol 50 no 10 pp1400ndash1409 2011

[157] D J Tuma ldquoRole of malondialdehyde-acetaldehyde adducts inliver injuryrdquo Free Radical Biology and Medicine vol 32 no 4pp 303ndash308 2002

[158] B P Sampey S Korourian M J Ronis T M Badger and DR Petersen ldquoImmunohistochemical characterization of hepaticmalondialdehyde and 4-hydroxynonenal modified proteinsduring early stages of ethanol-induced liver injuryrdquo AlcoholismClinical and Experimental Research vol 27 no 6 pp 1015ndash10222003

[159] E Albano ldquoRole of adaptive immunity in alcoholic liverdiseaserdquo International Journal of Hepatology vol 2012 ArticleID 893026 7 pages 2012

[160] G M Thiele L W Klassen and D J Tuma ldquoFormation andimmunological properties of aldehyde-derived protein adductsfollowing alcohol consumptionrdquoMethods in Molecular Biologyvol 447 pp 235ndash257 2008

[161] S K Das and D M Vasudevan ldquoAlcohol-induced oxidativestressrdquo Life Sciences vol 81 no 3 pp 177ndash187 2007

[162] O Niemela ldquoDistribution of ethanol-induced protein adductsin vivo relationship to tissue injuryrdquo Free Radical Biology andMedicine vol 31 no 12 pp 1533ndash1538 2001

[163] E Mottaran S F Stewart R Rolla et al ldquoLipid peroxidationcontributes to immune reactions associated with alcoholic liverdiseaserdquo Free Radical Biology and Medicine vol 32 no 1 pp38ndash45 2002

[164] M S Willis L W Klassen D J Tuma M F Sorrelland G M Thiele ldquoAdduction of soluble proteins withmalondialdehyde-acetaldehyde (MAA) induces antibody pro-duction and enhances T-cell proliferationrdquo Alcoholism Clinicaland Experimental Research vol 26 no 1 pp 94ndash106 2002


[165] X Dou S Li Z Wang et al ldquoInhibition of NF-120581B activation by4-hydroxynonenal contributes to liver injury in a mouse modelof alcoholic liver diseaserdquo The American Journal of Pathologyvol 181 no 5 pp 1702ndash1710 2012

[166] R L Smathers J J Galligan B J Stewart and D R PetersenldquoOverview of lipid peroxidation products and hepatic proteinmodification in alcoholic liver diseaserdquo Chemico-BiologicalInteractions vol 192 no 1-2 pp 107ndash112 2011

[167] G Poli F Biasi and G Leonarduzzi ldquo4-Hydroxynonenal-protein adducts a reliable biomarker of lipid oxidation in liverdiseasesrdquoMolecular Aspects of Medicine vol 29 no 1-2 pp 67ndash71 2008

[168] D R Petersen and J A Doorn ldquoReactions of 4-hydroxynonenalwith proteins and cellular targetsrdquo Free Radical Biology andMedicine vol 37 no 7 pp 937ndash945 2004

[169] J J Galligan R L Smathers K S Fritz L E Epperson L EHunter and D R Petersen ldquoProtein carbonylation in a murinemodel for early alcoholic liver diseaserdquo Chemical Research inToxicology vol 25 no 5 pp 1012ndash1021 2012

[170] M Perluigi R Coccia and D A Butterfield ldquo4-Hydroxy-2-nonenal a reactive product of lipid peroxidation and neurode-generative diseases a toxic combination illuminated by redoxproteomics studiesrdquo Antioxidants amp Redox Signaling vol 17 no11 pp 1590ndash1609 2012

[171] T T Reed ldquoLipid peroxidation and neurodegenerative diseaserdquoFree Radical Biology and Medicine vol 51 no 7 pp 1302ndash13192011

[172] K Zarkovic ldquo4-hydroxynonenal and neurodegenerative dis-easesrdquo Molecular Aspects of Medicine vol 24 no 4-5 pp 293ndash303 2003

[173] M L Selley ldquo(E)-4-hydroxy-2-nonenal may be involved in thepathogenesis of Parkinsonrsquos diseaserdquo Free Radical Biology andMedicine vol 25 no 2 pp 169ndash174 1998

[174] A Yoritaka N Hattori K Uchida M Tanaka E R Stadt-man and Y Mizuno ldquoImmunohistochemical detection of 4-hydroxynonenal protein adducts in Parkinson diseaserdquo Pro-ceedings of the National Academy of Sciences of the United Statesof America vol 93 no 7 pp 2696ndash2701 1996

[175] N Traverso S Menini E P Maineri et al ldquoMalondialdehyde alipoperoxidation-derived aldehyde can bring about secondaryoxidative damage to proteinsrdquo Journals of Gerontology A Bio-logical Sciences andMedical Sciences vol 59 no 9 pp 890ndash8952004

[176] D J Tuma M L Kearley G M Thiele et al ldquoElucidation ofreaction scheme describing malondialdehydemdashacetaldehydemdashprotein adduct formationrdquoChemical Research in Toxicology vol14 no 7 pp 822ndash832 2001

[177] G Wang H Li and M Firoze Khan ldquoDifferential oxida-tive modification of proteins in MRL++ and MRLlpr miceincreased formation of lipid peroxidation-derived aldehyde-protein adducts may contribute to accelerated onset of autoim-mune responserdquo Free Radical Research vol 46 no 12 pp 1472ndash1481 2012

[178] M J Duryee L W Klassen B L Jones M S Willis DJ Tuma and G M Thiele ldquoIncreased immunogenicity toP815 cells modified with malondialdehyde and acetaldehyderdquoInternational Immunopharmacology vol 8 no 8 pp 1112ndash11182008

[179] G Wang G A S Ansari and M F Khan ldquoInvolvement oflipid peroxidation-derived aldehyde-protein adducts in autoim-munity mediated by trichloroethenerdquo Journal of Toxicology and

Environmental Health A Current Issues vol 70 no 23 pp 1977ndash1985 2007

[180] M Wallberg J Bergquist A Achour E Breij and R AHarris ldquoMalondialdehyde modification of myelin oligoden-drocyte glycoprotein leads to increased immunogenicity andencephalitogenicityrdquo European Journal of Immunology vol 37no 7 pp 1986ndash1995 2007

[181] DWeismann and C J Binder ldquoThe innate immune response toproducts of phospholipid peroxidationrdquo Biochimica et Biophys-ica Acta Biomembranes vol 1818 no 10 pp 2465ndash2475 2012

[182] D A Slatter N C Avery and A J Bailey ldquoIdentification of anew cross-link and unique histidine adduct from bovine serumalbumin incubated withmalondialdehyderdquo Journal of BiologicalChemistry vol 279 no 1 pp 61ndash69 2004

[183] J Cheng F Wang D-F Yu P-F Wu and J-G Chen ldquoThecytotoxic mechanism of malondialdehyde and protective effectof carnosine via protein cross-linkingmitochondrial dysfunc-tionreactive oxygen speciesMAPKpathway in neuronsrdquoEuro-pean Journal of Pharmacology vol 650 no 1 pp 184ndash194 2011

[184] D Weismann K Hartvigsen N Lauer et al ldquoComplementfactor H binds malondialdehyde epitopes and protects fromoxidative stressrdquo Nature vol 478 no 7367 pp 76ndash81 2011

[185] M Veneskoski S P Turunen O Kummu et al ldquoSpecific recog-nition of malondialdehyde and malondialdehyde acetaldehydeadducts on oxidized LDL and apoptotic cells by complementanaphylatoxin C3ardquo Free Radical Biology and Medicine vol 51no 4 pp 834ndash843 2011

[186] K K Kharbanda K A Shubert T A Wyatt M F Sorrelland D J Tuma ldquoEffect of malondialdehyde-acetaldehyde-protein adducts on the protein kinase C-dependent secretion ofurokinase-type plasminogen activator in hepatic stellate cellsrdquoBiochemical Pharmacology vol 63 no 3 pp 553ndash562 2002

[187] L J Marnett ldquoOxy radicals lipid peroxidation and DNAdamagerdquo Toxicology vol 181-182 pp 219ndash222 2002

[188] L J Marnett ldquoLipid peroxidation-DNA damage by malondi-aldehyderdquoMutation Research vol 424 no 1-2 pp 83ndash95 1999

[189] S P Fink G R Reddy and L J Marnett ldquoMutagenicityin Escherichia coli of the major DNA adduct derived fromthe endogenous mutagen malondialdehyderdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 94 no 16 pp 8652ndash8657 1997

[190] M-L Vohringer T W Becker G Krieger H Jacobi andI Witte ldquoSynergistic DNA damaging effects of malondialde-hydeCu(II) in PM2DNA and in human fibroblastsrdquoToxicologyLetters vol 94 no 3 pp 159ndash166 1998

[191] C Ji C A Rouzer L J Marnett and J A Pietenpol ldquoInductionof cell cycle arrest by the endogenous product of lipid peroxida-tion malondialdehyderdquo Carcinogenesis vol 19 no 7 pp 1275ndash1283 1998

[192] M S Willis L W Klassen D L Carlson C F Brouse and GM Thiele ldquoMalondialdehyde-acetaldehyde haptenated proteinbindsmacrophage scavenger receptor(s) and induces lysosomaldamagerdquo International Immunopharmacology vol 4 no 7 pp885ndash899 2004

[193] M B Otteneder C G Knutson J S Daniels et al ldquoIn vivooxidative metabolism of a major peroxidation-derived DNAadduct M1dGrdquo Proceedings of the National Academy of Sciencesof the United States of America vol 103 no 17 pp 6665ndash66692006

[194] S D Cline M F Lodeiro L J Marnett C E Cameron andJ J Arnold ldquoArrest of human mitochondrial RNA polymerase


transcription by the biological aldehyde adduct ofDNAM1dGrdquoNucleic Acids Research vol 38 no 21 pp 7546ndash7557 2010

[195] R J Sram P Farmer R Singh et al ldquoEffect of vitamin levelson biomarkers of exposure and oxidative damage-the EXPAHstudyrdquoMutation Research Genetic Toxicology and Environmen-tal Mutagenesis vol 672 no 2 pp 129ndash134 2009

[196] C M Spickett ldquoThe lipid peroxidation product 4-hydroxy-2-nonenal advances in chemistry and analysisrdquo Redox Biologyvol 1 no 1 pp 145ndash152 2013

[197] P V Usatyuk and V Natarajan ldquoHydroxyalkenals and oxidizedphospholipids modulation of endothelial cytoskeleton focaladhesion and adherens junction proteins in regulating endothe-lial barrier functionrdquo Microvascular Research vol 83 no 1 pp45ndash55 2012

[198] R Sharma A Sharma P Chaudhary et al ldquoRole of 4-hydroxynonenal in chemopreventive activities of sulforaphanerdquoFree Radical Biology and Medicine vol 52 no 11-12 pp 2177ndash2185 2012

[199] P Zimniak ldquoRelationship of electrophilic stress to agingrdquo FreeRadical Biology and Medicine vol 51 no 6 pp 1087ndash1105 2011

[200] K S Fritz and D R Petersen ldquoExploring the biology oflipid peroxidation-derived protein carbonylationrdquo ChemicalResearch in Toxicology vol 24 no 9 pp 1411ndash1419 2011

[201] D A Butterfield T Reed and R Sultana ldquoRoles of 3-nitrotyrosine- and 4-hydroxynonenal-modified brain proteinsin the progression and pathogenesis of Alzheimerrsquos diseaserdquoFree Radical Research vol 45 no 1 pp 59ndash72 2011

[202] L M Balogh and W M Atkins ldquoInteractions of glutathionetransferases with 4-hydroxynonenalrdquo Drug MetabolismReviews vol 43 no 2 pp 165ndash178 2011

[203] A J Klil-Drori and A Ariel ldquo15-Lipoxygenases in cancer adouble-edged swordrdquo Prostaglandins amp Other Lipid Mediatorsvol 106 pp 16ndash22 2013

[204] A R Brash W E Boeglin and M S Chang ldquoDiscoveryof a second 15S-lipoxygenase in humansrdquo Proceedings of theNational Academy of Sciences of the United States of Americavol 94 no 12 pp 6148ndash6152 1997

[205] I Ivanov D Heydeck K Hofheinz et al ldquoMolecular enzymol-ogy of lipoxygenasesrdquo Archives of Biochemistry and Biophysicsvol 503 no 2 pp 161ndash174 2010

[206] H Takamura and HW Gardner ldquoOxygenation of (3Z)-alkenalto (2E)-4-hydroxy-2-alkenal in soybean seed (Glycine max L)rdquoBiochimica et Biophysica Acta Lipids and Lipid Metabolism vol1303 no 2 pp 83ndash91 1996

[207] C Schneider K A Tallman N A Porter and A R BrashldquoTwo distinct pathways of formation of 4-hydroxynonenalMechanisms of nonenzymatic transformation of the 9- and 13-hydroperoxides of linoleic acid to 4-hydroxyalkenalsrdquo Journalof Biological Chemistry vol 276 no 24 pp 20831ndash20838 2001

[208] Y Riahi G Cohen O Shamni and S Sasson ldquoSignaling andcytotoxic functions of 4-hydroxyalkenalsrdquo American Journal ofPhysiology Endocrinology and Metabolism vol 299 no 6 ppE879ndashE886 2010

[209] S V K Mahipal J Subhashini M C Reddy et al ldquoEffect of15-lipoxygenase metabolites 15-(S)-HPETE and 15-(S)-HETEon chronic myelogenous leukemia cell line K-562 reactiveoxygen species (ROS) mediate caspase-dependent apoptosisrdquoBiochemical Pharmacology vol 74 no 2 pp 202ndash214 2007

[210] K A Kumar K M Arunasree K R Roy et al ldquoEffectsof (15S)-hydroperoxyeicosatetraenoic acid and (15S)-hydroxyeicosatetraenoic acid on the acute-lymphoblastic-leukaemia cell line Jurkat activation of the Fas-mediated death

pathwayrdquo Biotechnology and Applied Biochemistry vol 52 no2 pp 121ndash133 2009

[211] P M Eckl ldquoGenotoxicity of HNErdquo Molecular Aspects ofMedicine vol 24 no 4-5 pp 161ndash165 2003

[212] W Siems and T Grune ldquoIntracellular metabolism of 4-hydroxynonenalrdquoMolecular Aspects of Medicine vol 24 no 4-5 pp 167ndash175 2003

[213] J Alary F Gueraud and J-P Cravedi ldquoFate of 4-hydro-xynonenal in vivo disposition andmetabolic pathwaysrdquoMolec-ular Aspects of Medicine vol 24 no 4-5 pp 177ndash187 2003

[214] K E McElhanon C Bose R Sharma L Wu Y C Awasthiand S P Singh ldquoGsta4 null mouse embryonic fibroblasts exhibitenhanced sensitivity to oxidants role of 4-hydroxynonenal inoxidant toxicityrdquo Open Journal of Apoptosis vol 2 no 1 2013

[215] W Black Y Chen AMatsumoto et al ldquoMolecularmechanismsofALDH3A1-mediated cellular protection against 4-hydroxy-2-nonenalrdquo Free Radical Biology and Medicine vol 52 no 9 pp1937ndash1944 2012

[216] D Kong and V Kotraiah ldquoModulation of aldehyde dehy-drogenase activity affects (plusmn)-4-hydroxy-2E-nonenal (HNE)toxicity and HNE-protein adduct levels in PC12 cellsrdquo Journalof Molecular Neuroscience vol 47 no 3 pp 595ndash603 2012

[217] Y Huang W Li and A N T Kong ldquoAnti-oxidative stress reg-ulator NF-E2-related factor 2 mediates the adaptive inductionof antioxidant and detoxifying enzymes by lipid peroxidationmetabolite 4-hydroxynonenalrdquo Cell amp Bioscience vol 2 no 1article 40 2012

[218] Y Zhang M Sano K Shinmura et al ldquo4-Hydroxy-2-nonenalprotects against cardiac ischemia-reperfusion injury via theNrf2-dependent pathwayrdquo Journal of Molecular and CellularCardiology vol 49 no 4 pp 576ndash586 2010

[219] R C M Siow T Ishii and G E Mann ldquoModulation of antiox-idant gene expression by 4-hydroxynonenal atheroprotectiverole of the Nrf2ARE transcription pathwayrdquo Redox Report vol12 no 1-2 pp 11ndash15 2007

[220] M Tanito M-P Agbaga and R E Anderson ldquoUpregulationof thioredoxin system via Nrf2-antioxidant responsive elementpathway in adaptive-retinal neuroprotection in vivo and invitrordquo Free Radical Biology and Medicine vol 42 no 12 pp1838ndash1850 2007

[221] T Ishii K Itoh E Ruiz et al ldquoRole of Nrf2 in the regulationof CD36 and stress protein expression in murine macrophagesactivation by oxidativelymodified LDL and 4-hydroxynonenalrdquoCirculation Research vol 94 no 5 pp 609ndash616 2004

[222] D M Miller I N Singh J A Wang and E D Hall ldquoAdminis-tration of the Nrf2-ARE activators sulforaphane and carnosicacid attenuates 4-hydroxy-2-nonenal-induced mitochondrialdysfunction ex vivordquo Free Radical Biology and Medicine vol 57pp 1ndash9 2013

[223] L Gan and J A Johnson ldquoOxidative damage and the Nrf2-ARE pathway in neurodegenerative diseasesrdquo Biochimica etBiophysica Acta Molecular Basis of Disease 2013

[224] H K Na and Y J Surh ldquoOncogenic potential of Nrf2 and itsprincipal target protein heme oxygenase-1rdquo Free Radical Biologyand Medicine vol 67 pp 353ndash365 2014

[225] H A Seo and I K Lee ldquoThe role of Nrf2 adipocyte differen-tiation obesity and insulin resistancerdquo Oxidative Medicine andCellular Longevity vol 2013 Article ID 184598 7 pages 2013

[226] T BDeramaudt CDill andMBonay ldquoRegulation of oxidativestress by Nrf2 in the pathophysiology of infectious diseasesrdquoMedecine et Maladies Infectieuses vol 43 no 3 pp 100ndash1072013


[227] A Grochot-Przeczek J Dulak and A Jozkowicz ldquoHaemoxygenase-1 non-canonical roles in physiology and pathologyrdquoClinical Science vol 122 no 3 pp 93ndash103 2012

[228] MH Lin J H Yen C YWeng LWang C LHa andM JWuldquoLipid peroxidation end product 4-hydroxy-trans-2-nonenaltriggers unfolded protein response and heme oxygenase-1expression in PC12 cells roles of ROS and MAPK pathwaysrdquoToxicology vol 315 pp 24ndash37 2014

[229] A Ishikado Y Nishio K Morino et al ldquoLow concentrationof 4-hydroxy hexenal increases heme oxygenase-1 expressionthrough activation of Nrf2 and antioxidative activity in vascularendothelial cellsrdquo Biochemical and Biophysical Research Com-munications vol 402 no 1 pp 99ndash104 2010

[230] K Ueda T Ueyama K-I Yoshida et al ldquoAdaptive HNE-Nrf2-HO-1 pathway against oxidative stress is associated withacute gastric mucosal lesionsrdquo American Journal of PhysiologyGastrointestinal and Liver Physiology vol 295 no 3 pp G460ndashG469 2008

[231] A Holmgren and J Lu ldquoThioredoxin and thioredoxin reduc-tase current research with special reference to human diseaserdquoBiochemical andBiophysical ResearchCommunications vol 396no 1 pp 120ndash124 2010

[232] Z-H Chen Y Saito Y Yoshida A Sekine N Noguchi andE Niki ldquo4-hydroxynonenal induces adaptive response andenhances PC12 cell tolerance primarily through induction ofthioredoxin reductase 1 via activation of Nrf2rdquo Journal ofBiological Chemistry vol 280 no 51 pp 41921ndash41927 2005

[233] S C Lu ldquoGlutathione synthesisrdquo Biochimica et Biophysica Actavol 1830 no 5 pp 3143ndash3153 2013

[234] C C FranklinD S Backos IMohar C CWhiteH J Formanand T J Kavanagh ldquoStructure function and post-translationalregulation of the catalytic and modifier subunits of glutamatecysteine ligaserdquo Molecular Aspects of Medicine vol 30 no 1-2pp 86ndash98 2009

[235] D S Backos K S Fritz J R Roede D R Petersen and CC Franklin ldquoPosttranslational modification and regulation ofglutamate-cysteine ligase by the 120572120573-unsaturated aldehyde 4-hydroxy-2-nonenalrdquo Free Radical Biology andMedicine vol 50no 1 pp 14ndash26 2011

[236] H Zhang A Shih A Rinna andH J Forman ldquoResveratrol and4-hydroxynonenal act in concert to increase glutamate cysteineligase expression and glutathione in human bronchial epithelialcellsrdquoArchives of Biochemistry and Biophysics vol 481 no 1 pp110ndash115 2009

[237] H Zhang N Court and H J Forman ldquoSubmicromolarconcentrations of 4-hydroxynonenal induce glutamate cysteineligase expression in HBE1 cellsrdquo Redox Report vol 12 no 1-2pp 101ndash106 2007

[238] K E Iles and R-M Liu ldquoMechanisms of Glutamate CysteineLigase (GCL) induction by 4-hydroxynonenalrdquo Free RadicalBiology and Medicine vol 38 no 5 pp 547ndash556 2005

[239] H J Forman D A Dickinson and K E Iles ldquoHNEmdashsignalingpathways leading to its eliminationrdquo Molecular Aspects ofMedicine vol 24 no 4-5 pp 189ndash194 2003

[240] E K Braithwaite M D Mattie and J H Freedman ldquoActivationofmetallothionein transcription by 4-hydroxynonenalrdquo Journalof Biochemical and Molecular Toxicology vol 24 no 5 pp 330ndash334 2010

[241] J F Reichard and D R Petersen ldquoHepatic stellate cells lack AP-1 responsiveness to electrophiles and phorbol 12-myristate-13-acetaterdquoBiochemical and Biophysical Research Communicationsvol 322 no 3 pp 842ndash853 2004

[242] K Kikuta A Masamune M Satoh N Suzuki and TShimosegawa ldquo4-Hydroxy-2 3-nonenal activates activatorprotein-1 and mitogen-activated protein kinases in rat pancre-atic stellate cellsrdquoWorld Journal of Gastroenterology vol 10 no16 pp 2344ndash2351 2004

[243] S Camandola G Poli and M P Mattson ldquoThe lipid peroxi-dation product 4-hydroxy-23-nonenal increases AP-1- bindingactivity through caspase activation in neuronsrdquo Journal ofNeurochemistry vol 74 no 1 pp 159ndash168 2000

[244] E Shaulian and M Karin ldquoAP-1 as a regulator of cell life anddeathrdquo Nature Cell Biology vol 4 no 5 pp E131ndashE136 2002

[245] E Shaulian ldquoAP-1mdashthe Jun proteins oncogenes or tumorsuppressors in disguiserdquo Cellular Signalling vol 22 no 6 pp894ndash899 2010

[246] M J Morgan and Z Liu ldquoCrosstalk of reactive oxygen speciesand NF-120581B signalingrdquo Cell Research vol 21 no 1 pp 103ndash1152011

[247] A Siomek ldquoNF-120581B signaling pathway and free radical impactrdquoActa Biochimica Polonica vol 59 no 3 pp 323ndash331 2012

[248] J H Lim J-C Lee Y H Lee et al ldquoSimvastatin preventsoxygen and glucose deprivationreoxygenation-induced deathof cortical neurons by reducing the production and toxicity of4-hydroxy-2E-nonenalrdquo Journal of Neurochemistry vol 97 no1 pp 140ndash150 2006

[249] K Kaarniranta T Ryhanen H M Karjalainen et al ldquoGel-danamycin increases 4-hydroxynonenal (HNE)-induced celldeath in human retinal pigment epithelial cellsrdquo NeuroscienceLetters vol 382 no 1-2 pp 185ndash190 2005

[250] S W Luckey M Taylor B P Sampey R I Scheinman andD R Petersen ldquo4-Hydroxynonenal decreases interleukin-6expression and protein production in primary rat Kupffercells by inhibiting nuclear factor-120581B activationrdquo Journal ofPharmacology and ExperimentalTherapeutics vol 302 no 1 pp296ndash303 2002

[251] HMinekura T Kumagai Y Kawamoto F Nara andKUchidaldquo4-Hydroxy-2-nonenal is a powerful endogenous inhibitor ofendothelial responserdquo Biochemical and Biophysical ResearchCommunications vol 282 no 2 pp 557ndash561 2001

[252] C Ji K R Kozak and L JMarnett ldquoI120581Bkinase amolecular tar-get for inhibition by 4-hydroxy-2-nonenalrdquo Journal of BiologicalChemistry vol 276 no 21 pp 18223ndash18228 2001

[253] S J Lee C E Kim K W Seo and C D Kim ldquoHNE-induced5-LO expression is regulated by NF-120581BERK and Sp1p38MAPK pathways via EGF receptor in murine macrophagesrdquoCardiovascular Research vol 88 no 2 pp 352ndash359 2010

[254] S J Lee K W Seo M R Yun et al ldquo4-hydroxynonenalenhances MMP-2 production in vascular smooth muscle cellsvia mitochondrial ROS-mediated activation of the AktNF-120581Bsignaling pathwaysrdquo Free Radical Biology and Medicine vol 45no 10 pp 1487ndash1492 2008

[255] H Raza A John E M Brown S Benedict and A Kam-bal ldquoAlterations in mitochondrial respiratory functions redoxmetabolism and apoptosis by oxidant 4-hydroxynonenal andantioxidants curcumin andmelatonin in PC12 cellsrdquo Toxicologyand Applied Pharmacology vol 226 no 2 pp 161ndash168 2008

[256] P E Malone and M R Hernandez ldquo4-Hydroxynonenal aproduct of oxidative stress leads to an antioxidant response inoptic nerve head astrocytesrdquo Experimental Eye Research vol 84no 3 pp 444ndash454 2007

[257] F Vaillancourt B Morquette Q Shi et al ldquoDifferential regu-lation of cyclooxygenase-2 and inducible nitric oxide synthase


by 4-hydroxynonenal in human osteoarthritic chondrocytesthrough ATF-2CREB-1 transactivation and concomitant inhi-bition of NF-120581B signaling cascaderdquo Journal of Cellular Biochem-istry vol 100 no 5 pp 1217ndash1231 2007

[258] HAmmaKNaruseN Ishiguro andM Sokabe ldquoInvolvementof reactive oxygen species in cyclic stretch-induced NF-120581Bactivation in human fibroblast cellsrdquo British Journal of Pharma-cology vol 145 no 3 pp 364ndash373 2005

[259] B Donath C Fischer S Page et al ldquoChlamydia pneumoniaeactivates IKKI120581B-mediated signaling which is inhibited by 4-HNE and following primary exposurerdquoAtherosclerosis vol 165no 1 pp 79ndash88 2002

[260] T Kim and Q Yang ldquoPeroxisome-proliferator-activated recep-tors regulate redox signaling in the cardiovascular systemrdquoWorld Journal of Cardiology vol 5 no 6 pp 164ndash174 2013

[261] M Ahmadian J M Suh N Hah et al ldquoPPAR120574 signaling andmetabolism the good the bad and the futurerdquoNatureMedicinevol 19 no 5 pp 557ndash566 2013

[262] G Barrera C Toaldo S Pizzimenti et al ldquoThe role of PPARligands in controlling growth-related gene expression and theirinteraction with lipoperoxidation productsrdquo PPAR Researchvol 2008 Article ID 524671 15 pages 2008

[263] Z Wang X Dou D Gu et al ldquo4-Hydroxynonenal differ-entially regulates adiponectin gene expression and secretionvia activating PPAR120574 and accelerating ubiquitin-proteasomedegradationrdquo Molecular and Cellular Endocrinology vol 349no 2 pp 222ndash231 2012

[264] S Pizzimenti S Laurora F Briatore C FerrettiMUDianzaniand G Barrera ldquoSynergistic effect of 4-hydroxynonenal andPPAR ligands in controlling human leukemic cell growth anddifferentiationrdquo Free Radical Biology and Medicine vol 32 no3 pp 233ndash245 2002

[265] A Cerbone C Toaldo S Laurora et al ldquo4-hydroxynonenaland PPAR120574 ligands affect proliferation differentiation andapoptosis in colon cancer cellsrdquo Free Radical Biology andMedicine vol 42 no 11 pp 1661ndash1670 2007

[266] M Almeida E Ambrogini L Han S C Manolagas andR L Jilka ldquoIncreased lipid oxidation causes oxidative stressincreased peroxisome proliferator-activated receptor-120574 expres-sion and diminished pro-osteogenic Wnt signaling in theskeletonrdquo Journal of Biological Chemistry vol 284 no 40 pp27438ndash27448 2009

[267] J D Coleman K S Prabhu J T Thompson et al ldquoTheoxidative stress mediator 4-hydroxynonenal is an intracellu-lar agonist of the nuclear receptor peroxisome proliferator-activated receptor-120573120575 (PPAR120573120575)rdquo Free Radical Biology andMedicine vol 42 no 8 pp 1155ndash1164 2007

[268] R Zheng I Po V Mishin et al ldquoThe generation of 4-hydroxynonenal an electrophilic lipid peroxidation end prod-uct in rabbit cornea organ cultures treated with UVB light andnitrogen mustardrdquo Toxicology and Applied Pharmacology vol272 no 2 pp 345ndash355 2013

[269] R Zheng D E Heck V Mishin et al ldquoModulation ofkeratinocyte expression of antioxidants by 4-hydroxynonenala lipid peroxidation end productrdquo Toxicology and AppliedPharmacology vol 275 no 2 pp 113ndash121 2014

[270] K Uchida and T Kumagai ldquo4-Hydroxy-2-nonenal as a COX-2 inducerrdquo Molecular Aspects of Medicine vol 24 no 4-5 pp213ndash218 2003

[271] M Parola G Robino F Marra et al ldquoHNE interacts directlywith JNK isoforms in human hepatic stellate cellsrdquo Journal ofClinical Investigation vol 102 no 11 pp 1942ndash1950 1998

[272] A Rinna and H J Forman ldquoSHP-1 inhibition by 4-hydroxynonenal activates Jun N-terminal kinase and glutamatecysteine ligaserdquo American Journal of Respiratory Cell andMolecular Biology vol 39 no 1 pp 97ndash104 2008

[273] R-M Liu Z Borok and H J Forman ldquo4-Hydroxy-2-nonenalincreases 120574-glutamylcysteine synthetase gene expression inalveolar epithelial cellsrdquo American Journal of Respiratory Celland Molecular Biology vol 24 no 4 pp 499ndash505 2001

[274] D A Dickinson K E Iles N Watanabe et al ldquo4-Hydroxyno-nenal induces glutamate cysteine ligase through JNK in HBE1cellsrdquo Free Radical Biology and Medicine vol 33 no 7 pp 974ndash987 2002

[275] C Marantos V Mukaro J Ferrante C Hii and A Fer-rante ldquoInhibition of the lipopolysaccharide-induced stimula-tion of the members of the MAPK family in human mono-cytesmacrophages by 4-hydroxynonenal a product of oxidizedomega-6 fatty acidsrdquoAmerican Journal of Pathology vol 173 no4 pp 1057ndash1066 2008

[276] Q Shi F Vaillancourt V Cote et al ldquoAlterations of metabolicactivity in human osteoarthritic osteoblasts by lipid peroxida-tion end product 4-hydroxynonenalrdquo Arthritis Research andTherapy vol 8 no 6 article R159 2006

[277] P V Usatyuk N L Parinandi and V Natarajan ldquoRedox reg-ulation of 4-hydroxy-2-nonenal-mediated endothelial barrierdysfunction by focal adhesion adherens and tight junctionproteinsrdquo Journal of Biological Chemistry vol 281 no 46 pp35554ndash35566 2006

[278] N Shibata Y Kato Y Inose et al ldquo4-hydroxy-2-nonenalupregulates and phosphorylates cytosolic phospholipase A

2

in cultured Ra2 microglial cells via MAPK pathwaysrdquo Neu-ropathology vol 31 no 2 pp 122ndash128 2011

[279] M Verslegers K Lemmens I Van Hove and L MoonsldquoMatrix metalloproteinase-2 and -9 as promising benefactorsin development plasticity and repair of the nervous systemrdquoProgress in Neurobiology vol 105 pp 60ndash78 2013

[280] S J Lee C E Kim M R Yun et al ldquo4-Hydroxynonenalenhances MMP-9 production in murine macrophages via 5-lipoxygenase-mediated activation of ERK and p38 MAPKrdquoToxicology and Applied Pharmacology vol 242 no 2 pp 191ndash198 2010

[281] K W Seo S J Lee C E Kim et al ldquoParticipation of 5-lipoxygenase-derived LTB4 in 4-hydroxynonenal-enhancedMMP-2 production in vascular smooth muscle cellsrdquoAtherosclerosis vol 208 no 1 pp 56ndash61 2010

[282] B Morquette Q Shi P Lavigne P Ranger J C Fernandes andM Benderdour ldquoProduction of lipid peroxidation products inosteoarthritic tissues new evidence linking 4-hydroxynonenalto cartilage degradationrdquoArthritis and Rheumatism vol 54 no1 pp 271ndash281 2006

[283] I Hers E E Vincent and J M Tavare ldquoAkt signalling in healthand diseaserdquo Cell Signaling vol 23 no 10 pp 1515ndash1527 2011

[284] N Chalhoub and S J Baker ldquoPTEN and the PI3-kinase pathwayin cancerrdquoAnnual Review of Pathology vol 4 pp 127ndash150 2009

[285] C T Shearn K S Fritz P Reigan and D R PetersenldquoModification of Akt2 by 4-hydroxynonenal inhibits insulin-dependent Akt signaling in HepG2 cellsrdquo Biochemistry vol 50no 19 pp 3984ndash3996 2011

[286] C T Shearn R L Smathers D S Backos P Reigan D JOrlicky and D R Petersen ldquoIncreased carbonylation of thelipid phosphatase PTEN contributes to Akt2 activation in amurine model of early alcohol-induced steatosisrdquo Free RadicalBiology and Medicine vol 65 pp 680ndash692 2013


[287] C T Shearn P Reigan and D R Petersen ldquoInhibition ofHydrogen peroxide signaling by 4-hydroxynonenal due todifferential regulation of Akt1 andAkt2 contributes to decreasesin cell survival and proliferation in hepatocellular carcinomacellsrdquo Free Radical Biology and Medicine vol 53 no 1 pp 1ndash112012

[288] R Vatsyayan P Chaudhary A Sharma et al ldquoRole of 4-hydroxynonenal in epidermal growth factor receptor-mediatedsignaling in retinal pigment epithelial cellsrdquo Experimental EyeResearch vol 92 no 2 pp 147ndash154 2011

[289] S Turban and E Hajduch ldquoProtein kinase C isoforms media-tors of reactive lipid metabolites in the development of insulinresistancerdquo FEBS Letters vol 585 no 2 pp 269ndash274 2011

[290] M Maggiora and M A Rossi ldquoThe exocytosis induced in HL-60 cells by 4-hydroxynonenal a lipid peroxidation product isnot prevented by reduced glutathionerdquo Cell Biochemistry andFunction vol 24 no 1 pp 1ndash6 2006

[291] M Maggiora and M A Rossi ldquoExperimental researches onthe role of phosphoinositide-specific phospholipase C in 4-hydroxynonenal induced exocytosisrdquo Cell Biochemistry andFunction vol 21 no 2 pp 155ndash160 2003

[292] M A Rossi C Di Mauro and M U Dianzani ldquoExperimentalstudies on the mechanism of phospholipase C activation by thelipid peroxidation products 4-hydroxynonenal and 2-nonenalrdquoInternational Journal of Tissue Reactions vol 23 no 2 pp 45ndash50 2001

[293] M A Rossi C Di Mauro H Esterbauer F Fidale and MU Dianzani ldquoActivation of phosphoinositide-specific phospho-lipase C of rat neutrophils by the chemotactic aldehydes 4-hydroxy-23-trans-nonenal and 4-hydroxy-23-trans-octenalrdquoCell Biochemistry and Function vol 12 no 4 pp 275ndash280 1994

[294] E B de Oliveira-Junior J Bustamante P E Newburger andA Condino-Neto ldquoThe human NADPH oxidase primary andsecondary defects impairing the respiratory burst function andthe microbicidal ability of phagocytesrdquo Scandinavian Journal ofImmunology vol 73 no 5 pp 420ndash427 2011

[295] R S Harry L A Hiatt D W Kimmel et al ldquoMetabolic impactof 4-hydroxynonenal on macrophage-like RAW 2647 functionand activationrdquo Chemical Research in Toxicology vol 25 no 8pp 1643ndash1651 2012

[296] E Chiarpotto C Domenicotti D Paola et al ldquoRegulation of rathepatocyte protein kinase C 120573 isoenzymes by the lipid peroxi-dation product 4-hydroxy-23-nonenal a signaling pathway tomodulate vesicular transport of glycoproteinsrdquoHepatology vol29 no 5 pp 1565ndash1572 1999

[297] D Paola C Domenicotti M Nitti et al ldquoOxidative stressinduces increase in intracellular amyloid 120573-protein productionand selective activation of 120573I and 120573II PKCs in NT2 cellsrdquoBiochemical andBiophysical ResearchCommunications vol 268no 2 pp 642ndash646 2000

[298] U M Marinari M Nitti M A Pronzato and C DomenicottildquoRole of PKC-dependent pathways inHNE-induced cell proteintransport and secretionrdquoMolecular Aspects of Medicine vol 24no 4-5 pp 205ndash211 2003

[299] M Nitti C Domenicotti C DrsquoAbramo et al ldquoActivation ofPKC-120573 isoforms mediates HNE-induced MCP-1 release bymacrophagesrdquo Biochemical and Biophysical Research Commu-nications vol 294 no 3 pp 547ndash552 2002

[300] K V Ramana A A Fadl R Tammali A B M Reddy AK Chopra and S K Srivastava ldquoAldose reductase mediates

the lipopolysaccharide-induced release of inflammatory medi-ators in RAW2647 murine macrophagesrdquo Journal of BiologicalChemistry vol 281 no 44 pp 33019ndash33029 2006

[301] M Dodson V Darley-Usmar and J Zhang ldquoCellular metabolicand autophagic pathways traffic control by redox signalingrdquoFree Radical Biology and Medicine vol 63 pp 207ndash221 2013

[302] B G Hill P Haberzettl Y Ahmed S Srivastava and ABhatnagar ldquoUnsaturated lipid peroxidation-derived aldehydesactivate autophagy in vascular smooth-muscle cellsrdquo Biochemi-cal Journal vol 410 no 3 pp 525ndash534 2008

[303] P Haberzettl and B G Hill ldquoOxidized lipids activate autophagyin a JNK-dependent manner by stimulating the endoplasmicreticulum stress responserdquoRedox Biology vol 1 no 1 pp 56ndash642013

[304] M Dodson Q Liang M S Johnson et al ldquoInhibition of gly-colysis attenuates 4-hydroxynonenal-dependent autophagy andexacerbates apoptosis in differentiated SH-SY5Yneuroblastomacellsrdquo Autophagy vol 9 no 12 pp 1996ndash2008 2013

[305] T U Krohne N K Stratmann J Kopitz and F G HolzldquoEffects of lipid peroxidation products on lipofuscinogenesisand autophagy in human retinal pigment epithelial cellsrdquoExperimental Eye Research vol 90 no 3 pp 465ndash471 2010

[306] F Fyhrquist O Saijonmaa and T Strandberg ldquoThe roles ofsenescence and telomere shortening in cardiovascular diseaserdquoNature Reviews Cardiology vol 10 no 5 pp 274ndash283 2013

[307] P L Olive ldquoEndogenous DNA breaks gammaH2AX and therole of telomeresrdquo Aging vol 1 no 2 pp 154ndash156 2009

[308] C Gunes and K L Rudolph ldquoThe role of telomeres in stem cellsand cancerrdquo Cell vol 152 no 3 pp 390ndash393 2013

[309] S Arguelles AMachado andA Ayala ldquoAdduct formation of 4-hydroxynonenal andmalondialdehyde with elongation factor-2in vitro and in vivordquo Free Radical Biology and Medicine vol 47no 3 pp 324ndash330 2009

[310] C Wang M Maddick S Miwa et al ldquoAdult-onset short-termdietary restriction reduces cell senescence in micerdquo Aging vol2 no 9 pp 555ndash566 2010

[311] G Nelson J Wordsworth C Wang et al ldquoA senescent cellbystander effect senescence-induced senescencerdquo Aging Cellvol 11 no 2 pp 345ndash349 2012

[312] G Voghel N Thorin-Trescases N Farhat et al ldquoChronictreatment with N-acetyl-cystein delays cellular senescence inendothelial cells isolated from a subgroup of atheroscleroticpatientsrdquo Mechanisms of Ageing and Development vol 129 no5 pp 261ndash270 2008

[313] S Pizzimenti F Briatore S Laurora et al ldquo4-Hydroxynonenalinhibits telomerase activity and hTERT expression in humanleukemic cell linesrdquo Free Radical Biology and Medicine vol 40no 9 pp 1578ndash1591 2006

[314] S Pizzimenti E Menegatti D Berardi et al ldquo4-Hydroxynonenal a lipid peroxidation product of dietarypolyunsaturated fatty acids has anticarcinogenic properties incolon carcinoma cell lines through the inhibition of telomeraseactivityrdquo Journal of Nutritional Biochemistry vol 21 no 9 pp818ndash826 2010

[315] A Rufini P Tucci I Celardo and G Melino ldquoSenescence andaging the critical roles of p53rdquo Oncogene vol 32 no 43 pp5129ndash5143 2013

[316] Y Qian and X Chen ldquoSenescence regulation by the p53 proteinfamilyrdquoMethods in Molecular Biology vol 965 pp 37ndash61 2013

[317] E Sahin and R A DePinho ldquoAxis of ageing telomeres p53 andmitochondriardquo Nature Reviews Molecular Cell Biology vol 13no 6 pp 397ndash404 2012


[318] D Liu and Y Xu ldquoP53 oxidative stress and agingrdquoAntioxidantsand Redox Signaling vol 15 no 6 pp 1669ndash1678 2011

[319] H Hafsi and P Hainaut ldquoRedox control and interplay betweenp53 isoforms roles in the regulation of basal p53 levels cell fateand senescencerdquo Antioxidants and Redox Signaling vol 15 no6 pp 1655ndash1667 2011

[320] A Vigneron and K H Vousden ldquop53 ROS and senescence inthe control of agingrdquo Aging vol 2 no 8 pp 471ndash474 2010

[321] E H Verbon J A Post and J Boonstra ldquoThe influence ofreactive oxygen species on cell cycle progression inmammaliancellsrdquo Gene vol 511 no 1 pp 1ndash6 2012

[322] J Chiu and I W Dawes ldquoRedox control of cell proliferationrdquoTrends in Cell Biology vol 22 no 11 pp 592ndash601 2012

[323] S Lim and P Kaldis ldquoCdks cyclins and CKIs roles beyond cellcycle regulationrdquo Development vol 140 no 15 pp 3079ndash30932013

[324] G Barrera S Pizzimenti S Laurora EMoroni B Giglioni andM U Dianzani ldquo4-hydroxynonenal affects pRbE2F pathwayin HL-60 human leukemic cellsrdquo Biochemical and BiophysicalResearch Communications vol 295 no 2 pp 267ndash275 2002

[325] S Pizzimenti G Barrera M U Dianzani and S BrusselbachldquoInhibition of D1 D2 and A cyclin expression in HL-60 cells bythe lipid peroxydation product 4-hydroxynonenalrdquoFree RadicalBiology and Medicine vol 26 no 11-12 pp 1578ndash1586 1999

[326] O A Skorokhod L Caione T Marrocco et al ldquoInhibitionof erythropoiesis in malaria anemia role of hemozoin andhemozoin-generated 4-hydroxynonenalrdquo Blood vol 116 no 20pp 4328ndash4337 2010

[327] C D Albright E Klem A A Shah and P Gallagher ldquoBreastcancer cell-targeted oxidative stress enhancement of cancercell uptake of conjugated linoleic acid activation of p53and inhibition of proliferationrdquo Experimental and MolecularPathology vol 79 no 2 pp 118ndash125 2005

[328] S B Sunjic A Cipak F Rabuzin R Wildburger and NZarkovic ldquoThe influence of 4-hydroxy-2-nonenal on prolif-eration differentiation and apoptosis of human osteosarcomacellsrdquo BioFactors vol 24 no 1ndash4 pp 141ndash148 2005

[329] G Muzio A Trombetta G Martinasso R A Canuto and MMaggiora ldquoAntisense oligonucleotides against aldehyde dehy-drogenase 3 inhibit hepatoma cell proliferation by affectingMAP kinasesrdquo Chemico-Biological Interactions vol 143-144 pp37ndash43 2003

[330] R A Canuto G Muzio M Ferro et al ldquoInhibition of class-3 aldehyde dehydrogenase and cell growth by restored lipidperoxidation in hepatoma cell linesrdquo Free Radical Biology andMedicine vol 26 no 3-4 pp 333ndash340 1999

[331] S Pizzimenti G Barrera E Calzavara et al ldquoDown-regulationofNotch1 expression is involved inHL-60 cell growth inhibitioninduced by 4-hydroxynonenal a product of lipid peroxidationrdquoMedicinal Chemistry vol 4 no 6 pp 551ndash557 2008

[332] S Laurora E Tamagno F Briatore et al ldquo4-Hydroxynonenalmodulation of p53 family gene expression in the SK-N-BEneuroblastoma cell linerdquo Free Radical Biology andMedicine vol38 no 2 pp 215ndash225 2005

[333] G Barrera S Martinotti V Fazio et al ldquoEffect of 4-hydroxynonenal on c-myc expressionrdquo Toxicologic Pathologyvol 15 no 2 pp 238ndash240 1987

[334] G Barrera R Muraca S Pizzimenti et al ldquoInhibition of c-myc expression induced by 4-hydroxynonenal a product oflipid peroxidation in the HL-60 human leukemic cell linerdquoBiochemical and Biophysical Research Communications vol 203no 1 pp 553ndash561 1994

[335] M Rinaldi G Barrera P Spinsanti et al ldquoGrowth inhibitionand differentiation induction in murine erythroleukemia cellsby 4-hydroxynonenalrdquo Free Radical Research vol 34 no 6 pp629ndash637 2001

[336] G Barrera S Pizzimenti and M U Dianzani ldquo4-Hydroxy-nonenal and regulation of cell cycle effects on the pRbE2Fpathwayrdquo Free Radical Biology and Medicine vol 37 no 5 pp597ndash606 2004

[337] G Barrera S Pizzimenti R Muraca et al ldquoEffect of 4-hydroxynonenal on cell cycle progression and expression ofdifferentiation-associated antigens in HL-60 cellsrdquo Free RadicalBiology and Medicine vol 20 no 3 pp 455ndash462 1996

[338] P Chaudhary R Sharma M Sahu J K Vishwanatha SAwasthi and Y C Awasthi ldquo4-Hydroxynonenal induces G2Mphase cell cycle arrest by activation of the ataxia telangiectasiamutated and Rad3-related protein (ATR)checkpoint kinase 1(Chk1) signaling pathwayrdquo Journal of Biological Chemistry vol288 no 28 pp 20532ndash20546 2013

[339] XWang Y YangD RMoore S LNimmo S A Lightfoot andM M Huycke ldquo4-hydroxy-2-nonenal mediates genotoxicityand bystander effects caused by enterococcus faecalis-infectedmacrophagesrdquo Gastroenterology vol 142 no 3 pp 543ndash5512012

[340] P Pettazzoni S Pizzimenti C Toaldo et al ldquoInduction ofcell cycle arrest and DNA damage by the HDAC inhibitorpanobinostat (LBH589) and the lipid peroxidation end product4-hydroxynonenal in prostate cancer cellsrdquo Free Radical Biologyand Medicine vol 50 no 2 pp 313ndash322 2011

[341] Z F Peng C H V Koh Q T Li et al ldquoDeciphering themechanism of HNE-induced apoptosis in cultured murine cor-tical neurons transcriptional responses and cellular pathwaysrdquoNeuropharmacology vol 53 no 5 pp 687ndash698 2007

[342] T-J Lee J-T Lee S-K Moon C-H Kim J-W Park and TK Kwon ldquoAge-related differential growth rate and responseto 4-hydroxynonenal in mouse aortic smooth muscle cellsrdquoInternational Journal of Molecular Medicine vol 17 no 1 pp29ndash35 2006

[343] H Kakishita and Y Hattori ldquoVascular smooth muscle cellactivation and growth by 4-hydroxynonenalrdquo Life Sciences vol69 no 6 pp 689ndash697 2001

[344] R Tammali A Saxena S K Srivastava and K V RamanaldquoAldose reductase regulates vascular smooth muscle cell pro-liferation by modulating G1S phase transition of cell cyclerdquoEndocrinology vol 151 no 5 pp 2140ndash2150 2010

[345] C-D Huang H-H Chen C-H Wang et al ldquoHumanneutrophil-derived elastase induces airway smooth muscle cellproliferationrdquo Life Sciences vol 74 no 20 pp 2479ndash2492 2004

[346] S Pizzimenti C Toaldo P Pettazzoni M U Dianzani andG Barrera ldquoThe ldquotwo-facedrdquo effects of reactive oxygen speciesand the lipid peroxidation product 4-hydroxynonenal in thehallmarks of cancerrdquo Cancers vol 2 no 2 pp 338ndash363 2010

[347] D Trachootham J Alexandre and P Huang ldquoTargeting can-cer cells by ROS-mediated mechanisms a radical therapeuticapproachrdquo Nature Reviews Drug Discovery vol 8 no 7 pp579ndash591 2009

[348] H PelicanoD Carney and PHuang ldquoROS stress in cancer cellsand therapeutic implicationsrdquo Drug Resistance Updates vol 7no 2 pp 97ndash110 2004

[349] E O Hileman J Liu M Albitar M J Keating and P HuangldquoIntrinsic oxidative stress in cancer cells a biochemical basis fortherapeutic selectivityrdquo Cancer Chemotherapy and Pharmacol-ogy vol 53 no 3 pp 209ndash219 2004


[350] P Chaudhary R Sharma A Sharma et al ldquoMechanisms of 4-hydroxy-2-nonenal induced pro- and anti-apoptotic signalingrdquoBiochemistry vol 49 no 29 pp 6263ndash6275 2010

[351] R M Locksley N Killeen and M J Lenardo ldquoThe TNF andTNF receptor superfamilies integrating mammalian biologyrdquoCell vol 104 no 4 pp 487ndash501 2001

[352] S Elmore ldquoApoptosis a review of programmed cell deathrdquoToxicologic Pathology vol 35 no 4 pp 495ndash516 2007

[353] J L Franklin ldquoRedox regulation of the intrinsic pathway inneuronal apoptosisrdquo Antioxidants and Redox Signaling vol 14no 8 pp 1437ndash1448 2011

[354] S Haupt M Berger Z Goldberg and Y Haupt ldquoApoptosismdashthe p53 networkrdquo Journal of Cell Science vol 116 no 20 pp4077ndash4085 2003

[355] S O Abarikwu A B Pant and E O Farombi ldquo4-hydroxynonenal induces mitochondrial-mediated apoptosisand oxidative stress in SH-SY5Y human neuronal cellsrdquo Basicand Clinical Pharmacology and Toxicology vol 110 no 5 pp441ndash448 2012

[356] A Sharma R Sharma P Chaudhary et al ldquo4-hydroxynonenalinduces p53-mediated apoptosis in retinal pigment epithelialcellsrdquo Archives of Biochemistry and Biophysics vol 480 no 2pp 85ndash94 2008

[357] R Sharma A Sharma S Dwivedi P Zimniak S Awasthiand Y C Awasthi ldquo4-hydroxynonenal self-limits Fas-mediatedDISC-independent apoptosis by promoting export of Daxxfrom the nucleus to the cytosol and its binding to FasrdquoBiochemistry vol 47 no 1 pp 143ndash156 2008

[358] F Vaillancourt H Fahmi Q Shi et al ldquo4-hydroxynonenalinduces apoptosis in human osteoarthritic chondrocytes theprotective role of glutathione-S-transferaserdquo Arthritis ResearchandTherapy vol 10 no 5 article R107 2008

[359] Y C Awasthi R Sharma A Sharma et al ldquoSelf-regulatoryrole of 4-hydroxynonenal in signaling for stress-induced pro-grammed cell deathrdquo Free Radical Biology andMedicine vol 45no 2 pp 111ndash118 2008

[360] J A Doorn and D R Petersen ldquoCovalent modification ofamino acid nucleophiles by the lipid peroxidation products 4-hydroxy-2-nonenal and 4-oxo-2-nonenalrdquo Chemical Researchin Toxicology vol 15 no 11 pp 1445ndash1450 2002

[361] L M Sayre D Lin Q Yuan X Zhu and X Tang ldquoProteinadducts generated from products of lipid oxidation focus onHNE and ONErdquo Drug Metabolism Reviews vol 38 no 4 pp651ndash675 2006

[362] C A Monroy J A Doorn and D L Roman ldquoModificationand functional inhibition of regulator of G-protein signaling4 (RGS4) by 4-Hydroxy-2-nonenalrdquo Chemical Research inToxicology vol 26 no 12 pp 1832ndash1839 2013

[363] G Poli R J Schaur W G Siems and G Leonarduzzildquo4-hydroxynonenal a membrane lipid oxidation product ofmedicinal interestrdquo Medicinal Research Reviews vol 28 no 4pp 569ndash631 2008

[364] S Choudhury J Pan S Amin F-L Chung and R RoyldquoRepair kinetics of trans-4-Hydroxynonenal-induced cyclic1N2-propanodeoxyguanine DNA adducts by human cellnuclear extractsrdquo Biochemistry vol 43 no 23 pp 7514ndash75212004

[365] S Choudhury M Dyba J Pan R Roy and F L ChungldquoRepair kinetics of acrolein- and (E)-4-hydroxy-2-nonenal-derived DNA adducts in human colon cell extractsrdquo MutationResearch vol 751-752 pp 15ndash23 2013

[366] L Gros A A Ishchenko and M Saparbaev ldquoEnzymology ofrepair of etheno-adductsrdquoMutation Research Fundamental andMolecular Mechanisms of Mutagenesis vol 531 no 1-2 pp 219ndash229 2003

[367] J Nair P Srivatanakul C Haas et al ldquoHigh urinary excretionof lipid peroxidation-derived DNA damage in patients withcancer-prone liver diseasesrdquo Mutation Research Fundamentaland Molecular Mechanisms of Mutagenesis vol 683 no 1-2 pp23ndash28 2010

[368] J Nair F Gansauge H Beger P Dolara G Winde and HBartsch ldquoIncreased etheno-DNA adducts in affected tissues ofpatients suffering from Crohnrsquos disease ulcerative colitis andchronic pancreatitisrdquo Antioxidants and Redox Signaling vol 8no 5-6 pp 1003ndash1010 2006

[369] S Richard and J Lewis Hazardous Chemicals Desk ReferenceJohn Wiley amp Sons 2008

[370] A Ayala J Parrado M Bougria and A Machado ldquoEffect ofoxidative stress produced by cumene hydroperoxide on thevarious steps of protein synthesis Modifications of elongationfactor-2rdquo Journal of Biological Chemistry vol 271 no 38 pp23105ndash23110 1996

[371] J Parrado M Bougria A Ayala A Castano and A MachadoldquoEffects of aging on the various steps of protein synthesisfragmentation of elongation factor 2rdquo Free Radical Biology andMedicine vol 26 no 3-4 pp 362ndash370 1999

[372] J Parrado M Bougria A Ayala and A MacHado ldquoInducedmono-(ADP)-ribosylation of rat liver cytosolic proteins by lipidperoxidant agentsrdquo Free Radical Biology and Medicine vol 26no 9-10 pp 1079ndash1084 1999

[373] J Parrado E H Absi A Machado and A Ayala ldquolsquoIn vitrorsquoeffect of cumene hydroperoxide on hepatic elongation factor-2and its protection by melatoninrdquo Biochimica et Biophysica ActaGeneral Subjects vol 1624 no 1ndash3 pp 139ndash144 2003

[374] S Arguelles A Machado and A Ayala ldquolsquoIn vitrorsquo effect of lipidperoxidation metabolites on elongation factor-2rdquo Biochimica etBiophysica Acta General Subjects vol 1760 no 3 pp 445ndash4522006

[375] S Arguelles M Cano A Machado and A Ayala ldquoEffect ofaging and oxidative stress on elongation factor-2 in hypothala-mus and hypophysisrdquo Mechanisms of Ageing and Developmentvol 132 no 1-2 pp 55ndash64 2011

[376] S Arguelles M F Munoz M Cano A Machado and AAyala ldquoIn vitro and in vivo protection by melatonin againstthe decline of elongation factor-2 caused by lipid peroxidationpreservation of protein synthesisrdquo Journal of Pineal Researchvol 53 no 1 pp 1ndash10 2012

[377] S Arguelles A Machado and A Ayala ldquorsquoIn vitrorsquo protectiveeffect of a hydrophilic vitamin E analogue on the decrease inlevels of elongation factor 2 in conditions of oxidative stressrdquoGerontology vol 53 no 5 pp 282ndash288 2007

[378] S ArguellesM Cano AMachado andA Ayala ldquoComparativestudy of the In Vitro protective effects of several antioxidantson elongation factor 2 under oxidative stress conditionsrdquoBioscience Biotechnology and Biochemistry vol 74 no 7 pp1373ndash1379 2010

[379] S Arguelles S Camandola E R Hutchison R G Cutler AAyala and M P Mattson ldquoMolecular control of the amountsubcellular location and activity state of translation elongationfactor 2 in neurons experiencing stressrdquo Free Radical Biologyand Medicine vol 61 pp 61ndash71 2013

[380] S Arguelles S Camandola R G Cutler A Ayala and MP Mattson ldquoElongation factor 2 diphthamide is critical for


translation of two IRES-dependent protein targets XIAP andFGF2 under oxidative stress conditionsrdquo Free Radical Biologyand Medicine vol 67 pp 131ndash138 2013

[381] Y G Aboua N Brooks R Z Mahfouz A Agarwal and S S duPlessis ldquoA red palm oil diet can reduce the effects of oxidativestress on rat spermatozoardquo Andrologia vol 44 supplement 1pp 32ndash40 2012

[382] T R Kumar andMMuralidhara ldquoInduction of oxidative stressby organic hydroperoxides in testis and epididymal sperm ofrats in vivordquo Journal of Andrology vol 28 no 1 pp 77ndash85 2007

[383] T S ChanN Shangari J XWilsonHChan R F Butterworthand P J OrsquoBrien ldquoThe biosynthesis of ascorbate protectsisolated rat hepatocytes from cumene hydroperoxide-mediatedoxidative stressrdquo Free Radical Biology and Medicine vol 38 no7 pp 867ndash873 2005

[384] A A Shvedova E R Kisin A R Murray et al ldquoAntioxidantbalance and free radical generation in vitamin E-deficient miceafter dermal exposure to cumene hydroperoxiderdquo ChemicalResearch in Toxicology vol 15 no 11 pp 1451ndash1459 2002

[385] A Alam M Iqbal M Saleem S-U Ahmed and S Sul-tana ldquoMyrica nagi attenuates cumene hydroperoxide-inducedcutaneous oxidative stress and toxicity in Swiss albino micerdquoPharmacology and Toxicology vol 86 no 5 pp 209ndash214 2000

[386] M Jamal A Masood R Belcastro et al ldquoLipid hydroperoxideformation regulates postnatal rat lung cell apoptosis and alve-ologenesisrdquo Free Radical Biology and Medicine vol 55 pp 83ndash92 2013

[387] C O Hong C H Rhee N H Won H D Choi and K W LeeldquoProtective effect of 70 ethanolic extract of Lindera obtusilobaBlume on tert-butyl hydroperoxide-induced oxidative hepato-toxicity in ratsrdquo Food and Chemical Toxicology vol 53 pp 214ndash220 2013

[388] JMOh Y S Jung B S Jeon et al ldquoEvaluation of hepatotoxicityand oxidative stress in rats treated with tert-butyl hydroperox-iderdquo Food and Chemical Toxicology vol 50 no 5 pp 1215ndash12212012

[389] M-K Kim H-S Lee E-J Kim et al ldquoProtective effect of aque-ous extract of Perilla frutescens on tert-butyl hydroperoxide-induced oxidative hepatotoxicity in ratsrdquo Food and ChemicalToxicology vol 45 no 9 pp 1738ndash1744 2007

[390] P Kaur G Kaur and M P Bansal ldquoTertiary-butyl hydroper-oxide induced oxidative stress and male reproductive activityin mice role of transcription factor NF-120581B and testicularantioxidant enzymesrdquoReproductive Toxicology vol 22 no 3 pp479ndash484 2006

[391] C L Liu J M Wang C Y Chu M T Cheng and T H TsengldquoIn vivo protective effect of protocatechuic acid on tert-butylhydroperoxide-induced rat hepatotoxicityrdquo Food and ChemicalToxicology vol 40 no 5 pp 635ndash641 2002

[392] S Hix M B Kadiiska R P Mason and O Augusto ldquoIn vivometabolism of tert-Butyl hydroperoxide to methyl radicalsEPR spin-trapping and DNA methylation studiesrdquo ChemicalResearch in Toxicology vol 13 no 10 pp 1056ndash1064 2000

[393] J Q Ma J Ding L Zhang and C M Liu ldquoHepatoprotectiveproperties of sesamin against CCl4 induced oxidative stress-mediated apoptosis in mice via JNK pathwayrdquo Food andChemical Toxicology vol 64 pp 41ndash48 2013

[394] Y H Yeh Y L Hsieh and Y T Lee ldquoEffects of yam peelextract against carbon tetrachloride-induced hepatotoxicity inratsrdquo Journal of Agricultural and Food Chemistry vol 61 no 30pp 7387ndash7396 2013

[395] L Knockaert A Berson C Ribault et al ldquoCarbon tetra-chloride-mediated lipid peroxidation induces early mitochon-drial alterations in mouse liverrdquo Laboratory Investigation vol92 no 3 pp 396ndash410 2012

[396] J-H Choi D-W Kim N Yun et al ldquoProtective effects ofhyperoside against carbon tetrachloride-induced liver damagein micerdquo Journal of Natural Products vol 74 no 5 pp 1055ndash1060 2011

[397] H-Y Kim J-K Kim J-H Choi et al ldquoHepatoprotective effectof pinoresinol on carbon tetrachloride-induced hepatic damagein micerdquo Journal of Pharmacological Sciences vol 112 no 1 pp105ndash112 2010

[398] H Wang W Wei N-P Wang et al ldquoMelatonin amelioratescarbon tetrachloride-induced hepatic fibrogenesis in rats viainhibition of oxidative stressrdquo Life Sciences vol 77 no 15 pp1902ndash1915 2005

[399] R Lugo-Huitron P Ugalde Muniz B Pineda J Pedraza-Chaverrı C Rıos and V Perez-de la Cruz ldquoQuinolinic acidan endogenous neurotoxin with multiple targetsrdquo OxidativeMedicine and Cellular Longevity vol 2013 Article ID 10402414 pages 2013

[400] P D Maldonado V Perez-De La Cruz M Torres-Ramos et alldquoSelenium-induced antioxidant protection recruits modulationof thioredoxin reductase during excitotoxicpro-oxidant eventsin the rat striatumrdquo Neurochemistry International vol 61 no 2pp 195ndash206 2012

[401] S Sreekala and M Indira ldquoImpact of co administration ofselenium and quinolinic acid in the ratrsquos brainrdquo Brain Researchvol 1281 pp 101ndash107 2009

[402] J K Ryu H B Choi and J G McLarnon ldquoPeripheral benzodi-azepine receptor ligand PK11195 reduces microglial activationand neuronal death in quinolinic acid-injected rat striatumrdquoNeurobiology of Disease vol 20 no 2 pp 550ndash561 2005

[403] J I Rossato G Zeni C F Mello M A Rubin and J B TRocha ldquoEbselen blocks the quinolinic acid-induced productionof thiobarbituric acid reactive species but does not prevent thebehavioral alterations produced by intra-striatal quinolinic acidadministration in the ratrdquo Neuroscience Letters vol 318 no 3pp 137ndash140 2002

[404] A Santamarıa M E Jimenez-Capdeville A Camacho ERodrıguez-Martınez A Flores and S Galvan-Arzate ldquoIn vivohydroxyl radical formation after quinolinic acid infusion intorat corpus striatumrdquoNeuroReport vol 12 no 12 pp 2693ndash26962001

[405] E Rodrıguez-Martınez A Camacho P D Maldonado et alldquoEffect of quinolinic acid on endogenous antioxidants in ratcorpus striatumrdquo Brain Research vol 858 no 2 pp 436ndash4392000

[406] K Jomova andM Valko ldquoAdvances in metal-induced oxidativestress and human diseaserdquo Toxicology vol 283 no 2-3 pp 65ndash87 2011

[407] A Boveris R Musacco-Sebio N Ferrarotti et al ldquoThe acutetoxicity of iron and copper biomolecule oxidation and oxida-tive damage in rat liverrdquo Journal of Inorganic Biochemistry vol116 pp 63ndash69 2012

[408] D Ozcelik H Uzun and M Naziroglu ldquoN-acetylcysteineattenuates copper overload-induced oxidative injury in brain ofratrdquo Biological Trace Element Research vol 147 no 1ndash3 pp 292ndash298 2012

[409] A Alexandrova L Petrov A Georgieva et al ldquoEffect of copperintoxication on rat liver proteasome activity relationship with


oxidative stressrdquo Journal of Biochemical and Molecular Toxicol-ogy vol 22 no 5 pp 354ndash362 2008

[410] B Scharf and L D Trombetta ldquoThe effects of the wood preser-vative copper dimethyldithiocarbamate in the hippocampus ofmaternal and newborn Long-Evans ratsrdquoToxicology Letters vol174 no 1-3 pp 117ndash124 2007

[411] K Parveen M R Khan and W A Siddiqui ldquoPycnogenolprevents potassium dichromate (K2Cr2O7)-induced oxidativedamage and nephrotoxicity in ratsrdquo Chemico-Biological Interac-tions vol 181 no 3 pp 343ndash350 2009

[412] D Kotyzova A Hodkova M Bludovska and V Eybl ldquoEffectof chromium (VI) exposure on antioxidant defense statusand trace element homeostasis in acute experiment in ratrdquoToxicology and Industrial Health In press

[413] S Karaca and G Eraslan ldquoThe effects of flaxseed oil oncadmium-induced oxidative stress in ratsrdquo Biological TraceElement Research vol 155 no 3 pp 423ndash430 2013

[414] Q Chen R Zhang W Li et al ldquoThe protective effect ofgrape seed procyanidin extract against cadmium-induced renaloxidative damage in micerdquo Environmental Toxicology and Phar-macology vol 36 no 3 pp 759ndash768 2013

[415] P Leelavinothan and S Kalist ldquoBeneficial effect of hesperetinon cadmium induced oxidative stress in rats an in vivo and invitro studyrdquo European Review for Medical and PharmacologicalSciences vol 15 no 9 pp 992ndash1002 2011

[416] B I Ognjanovic S D Markovic N Z Ethordevic I STrbojevic A S Stajn and Z S Saicic ldquoCadmium-induced lipidperoxidation and changes in antioxidant defense system in therat testes protective role of coenzyme Q(10) and vitamin ErdquoReproductive Toxicology vol 29 no 2 pp 191ndash197 2010

[417] K Amudha and L Pari ldquoBeneficial role of naringin a flavanoidon nickel induced nephrotoxicity in ratsrdquo Chemico-BiologicalInteractions vol 193 no 1 pp 57ndash64 2011

[418] L Pari and K Amudha ldquoHepatoprotective role of naringin onnickel-induced toxicity in male Wistar ratsrdquo European Journalof Pharmacology vol 650 no 1 pp 364ndash370 2011

[419] A Scibior D Gołębiowska and I Niedzwiecka ldquoMagnesiumcan protect against vanadium-induced lipid peroxidation in thehepatic tissuerdquo Oxidative Medicine and Cellular Longevity vol2013 Article ID 802734 11 pages 2013

[420] A ScibiorH Zaporowska and INiedzwiecka ldquoLipid peroxida-tion in the kidney of rats treated with V andor Mg in drinkingwaterrdquo Journal of Applied Toxicology vol 30 no 5 pp 487ndash4962010

[421] A Scibior H Zaporowska J Ostrowski and A BanachldquoCombined effect of vanadium(V) and chromium(III) on lipidperoxidation in liver and kidney of ratsrdquo Chemico-BiologicalInteractions vol 159 no 3 pp 213ndash222 2006

[422] E N Martins N T C Pessano L Leal et al ldquoProtectiveeffect of Melissa officinalis aqueous extract against Mn-inducedoxidative stress in chronically exposed micerdquo Brain ResearchBulletin vol 87 no 1 pp 74ndash79 2012

[423] Y Chtourou H Fetoui M Sefi et al ldquoSilymarin a natu-ral antioxidant protects cerebral cortex against manganese-induced neurotoxicity in adult ratsrdquo BioMetals vol 23 no 6pp 985ndash996 2010

[424] M T Chen G W Cheng C C Lin B H Chen and YL Huang ldquoEffects of acute manganese chloride exposure onlipid peroxidation and alteration of trace metals in rat brainrdquoBiological Trace Element Research vol 110 no 2 pp 163ndash1782006

[425] S A Salama H A Omar I A Maghrabi M S Alsaeedand A E El-Tarras ldquoIron supplementation at high altitudesinduces inflammation and oxidative injury to lung tissues inratsrdquo Toxicology and Applied Pharmacology vol 274 no 1 pp1ndash6 2014

[426] J Kim H D Paik Y C Yoon and E Park ldquoWhey proteininhibits iron overload-induced oxidative stress in ratsrdquo Journalof Nutritional Science and Vitaminology vol 59 no 3 pp 198ndash205 2013

[427] L F Arruda S F ArrudaN A Campos F F deValencia and EM de Siqueira ldquoDietary iron concentrationmay influence agingprocess by altering oxidative stress in tissues of adult ratsrdquo PloSONE vol 8 no 4 Article ID e61058 2013

[428] H C Yu S F Feng P L Chao and A M Y Lin ldquoAnti-inflammatory effects of pioglitazone on iron-induced oxidativeinjury in the nigrostriatal dopaminergic systemrdquo Neuropathol-ogy and Applied Neurobiology vol 36 no 7 pp 612ndash622 2010

[429] S Oktar Z Yonden M Aydin S Ilhan E Alcin and O HOzturk ldquoProtective effects of caffeic acid phenethyl ester oniron-induced liver damage in ratsrdquo Journal of Physiology andBiochemistry vol 65 no 4 pp 339ndash344 2009

[430] A Kokoszko J Dabrowski A Lewinski and M Karbownik-Lewinska ldquoProtective effects of GH and IGF-I against iron-induced lipid peroxidation in vivordquo Experimental and Toxico-logic Pathology vol 60 no 6 pp 453ndash458 2008

[431] D S Maharaj H Maharaj S Daya and B D Glass ldquoMelatoninand 6-hydroxymelatonin protect against iron-induced neuro-toxicityrdquo Journal of Neurochemistry vol 96 no 1 pp 78ndash812006

[432] N P Morales Y Yamaguchi K Murakami N Kosem and HUtsumi ldquoHepatic reduction of carbamoyl-PROXYL in ferricnitrilotriacetate induced iron overloaded mice an in vivo ESRstudyrdquo Biological and Pharmaceutical Bulletin vol 35 no 7 pp1035ndash1040 2012

[433] W Volkel R Alvarez-Sanchez I Weick A Mally W Dekantand A Pahler ldquoGlutathione conjugates of 4-hydroxy-2(E)-nonenal as biomarkers of hepatic oxidative stress-induced lipidperoxidation in ratsrdquo Free Radical Biology andMedicine vol 38no 11 pp 1526ndash1536 2005

[434] V Eybl D Kotyzova P Cerna and J Koutensky ldquoEffect ofmelatonin curcumin quercetin and resveratrol on acute ferricnitrilotriacetate (Fe-NTA)-induced renal oxidative damage inratrdquoHuman and Experimental Toxicology vol 27 no 4 pp 347ndash353 2008

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


include (i) two derived from the phosphatidylinositol phos-phates diacylglycerol (DAG) and inositol phosphates (IPs)DAG is a physiological activator of protein kinase C [6 7]and transcription factor nuclear factor-kB (NF-120581B) whichpromotes cell survival and proliferation Diacylglycerol alsointeracts indirectly with other signalling molecules such assmall G proteins [8] IPs are a highly charged family oflipid-derived metabolites involved in signal transductionthat results in activation of Akt mTOR [9] and calcium-homeostasis [10 11] (ii) sphingosine-1-phosphate a sph-ingolipid derived from ceramide that is a potent messen-ger molecule involved in regulating calcium mobilizationmigration adhesion and proliferation [12ndash14] (iii) theprostaglandins which are one type of fatty-acid derivedeicosanoid involved in inflammation [15 16] and immunity[17] (iv) phosphatidylserine a phospholipid that plays animportant role in a number of signaling pathways includeskinases small GTPases and fusogenic proteins [18] (v) thesteroid hormones such as estrogen testosterone and cortisolwhich modulate a host of functions such as reproductionmetabolism stress response inflammation blood pressureand salt and water balance [19]

2 Lipids Damage by Reactive Oxygen Species

One of the consequences of uncontrolled oxidative stress(imbalance between the prooxidant and antioxidant levelsin favor of prooxidants) is cells tissues and organs injurycaused by oxidative damage It has long been recognized thathigh levels of free radicals or reactive oxygen species (ROS)can inflict direct damage to lipids The primary sources ofendogenous ROS production are the mitochondria plasmamembrane endoplasmic reticulum and peroxisomes [20]through a variety of mechanisms including enzymatic reac-tions andor autooxidation of several compounds suchas catecholamines and hydroquinone Different exogenousstimuli such as the ionizing radiation ultraviolet raystobacco smoke pathogen infections environmental toxinsand exposure to herbicideinsecticides are sources of in vivoROS production

The two most prevalent ROS that can affect profoundlythe lipids are mainly hydroxyl radical (HO∙) and hydroper-oxyl (HO∙

2

) The hydroxyl radical (HO∙) is a small highlymobile water-soluble and chemically most reactive speciesof activated oxygen This short-lived molecule can be pro-duced from O

2

in cell metabolism and under a variety ofstress conditions A cell produces around 50 hydroxyl radicalsevery second In a full day each cell would generate 4million hydroxyl radicals which can be neutralized or attackbiomolecules [21] Hydroxyl radicals cause oxidative damageto cells because they unspecifically attack biomolecules [22]located less than a few nanometres from its site of generationand are involved in cellular disorders such as neurodegenera-tion [23 24] cardiovascular disease [25] and cancer [26 27]It is generally assumed that HO∙ in biological systems isformed through redox cycling by Fenton reaction where freeiron (Fe2+) reacts with hydrogen peroxide (H

2

O2

) and theHaber-Weiss reaction that results in the production of Fe2+

Haber-Weiss reaction

Fenton reaction

O2

∙OH + H2OH2O2

H+

H+ M(n+1) Mn

O2∙minus

HO∙2

Figure 1 Fenton and Haber-Weiss reaction Reduced form oftransition-metals (M119899) reacts trough the Fenton reaction withhydrogen peroxide (H

2

O2

) leading to the generation of ∙OHSuperoxide radical (O

2

∙minus) can also react with oxidized form oftransitionmetals (M(119899+1)) in the Haber-Weiss reaction leading to theproduction of M119899 which then again affects redox cycling

when superoxide reacts with ferric iron (Fe3+) In additionto the iron redox cycling described above also a number ofother transition-metal including Cu Ni Co and V can beresponsible for HO∙ formation in living cells (Figure 1)

The hydroperoxyl radical (HO∙2

) plays an importantrole in the chemistry of lipid peroxidation This protonatedform of superoxide yields H

2

O2

which can react with redoxactive metals including iron or copper to further generateHO∙ through Fenton or Haber-Weiss reactions The HO∙

2

is a much stronger oxidant than superoxide anion-radicaland could initiate the chain oxidation of polyunsaturatedphospholipids thus leading to impairment of membranefunction [28ndash30]

21 Lipid Peroxidation Process Lipid peroxidation can bedescribed generally as a process under which oxidants suchas free radicals or nonradical species attack lipids containingcarbon-carbon double bond(s) especially polyunsaturatedfatty acids (PUFAs) that involve hydrogen abstraction froma carbon with oxygen insertion resulting in lipid per-oxyl radicals and hydroperoxides as described previously[31] Glycolipids phospholipids (PLs) and cholesterol (Ch)are also well-known targets of damaging and potentiallylethal peroxidative modification Lipids also can be oxi-dized by enzymes like lipoxygenases cyclooxygenases andcytochrome P450 (see above lipid as signaling molecules)In response to membrane lipid peroxidation and accord-ing to specific cellular metabolic circumstances and repaircapacities the cells may promote cell survival or inducecell death Under physiological or low lipid peroxidationrates (subtoxic conditions) the cells stimulate their mainte-nance and survival through constitutive antioxidants defensesystems or signaling pathways activation that upregulateantioxidants proteins resulting in an adaptive stress responseBy contrast under medium or high lipid peroxidation rates(toxic conditions) the extent of oxidative damage overwhelmsrepair capacity and the cells induce apoptosis or necrosisprogrammed cell death both processes eventually lead tomolecular cell damage which may facilitate development ofvarious pathological states and accelerated agingThe impactof lipids oxidation in cell membrane and how these oxidativedamages are involved in both physiological processes andmajor pathological conditions have been analysed in severalreviews [32ndash35]










2

O2



OOH2 3

1

Rearrangement

Unsaturated lipid

Unsaturated lipid



Lipid hydroperoxide

Antioxidant4

O2

OO∙

R∙ H+





LOOH +M119899+1 997888rarr LOO∙ +H+ +M119899 (2)



119892

1O2







2


2


2

)[4 91 92] PGH

2





2

and H2







[104ndash113][81 108 114ndash121]

Cancer MDA4-HNE



[72 79 109 123 135 137ndash141][72 104 109 131 135 138 139 142ndash144]

Diabetes MDA4-HNE

[79 109 123 140 145ndash150][131 135 142 143 151ndash156]




[81 108 114ndash121][72 114 131 135 142 170ndash174]

PUFAAA



Monocyclicperoxide

HHTMDA

Malonicsemialdehyde




1

2

3 33

4

546

7

Cyclization

CO2 + H2O

O2

O2

+ H+

H+

H+


Radical∙

PUFA-radical∙

2O2

PGG2

PGH2

TXA2


2

(TXA2












LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

























2

O2



OOH2 3

1

Rearrangement

Unsaturated lipid

Unsaturated lipid



Lipid hydroperoxide

Antioxidant4

O2

OO∙

R∙ H+





LOOH +M119899+1 997888rarr LOO∙ +H+ +M119899 (2)



119892

1O2







2


2


2

)[4 91 92] PGH

2





2

and H2







[104ndash113][81 108 114ndash121]

Cancer MDA4-HNE



[72 79 109 123 135 137ndash141][72 104 109 131 135 138 139 142ndash144]

Diabetes MDA4-HNE

[79 109 123 140 145ndash150][131 135 142 143 151ndash156]




[81 108 114ndash121][72 114 131 135 142 170ndash174]

PUFAAA



Monocyclicperoxide

HHTMDA

Malonicsemialdehyde




1

2

3 33

4

546

7

Cyclization

CO2 + H2O

O2

O2

+ H+

H+

H+


Radical∙

PUFA-radical∙

2O2

PGG2

PGH2

TXA2


2

(TXA2












LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















OOH2 3

1

Rearrangement

Unsaturated lipid

Unsaturated lipid



Lipid hydroperoxide

Antioxidant4

O2

OO∙

R∙ H+





LOOH +M119899+1 997888rarr LOO∙ +H+ +M119899 (2)



119892

1O2







2


2


2

)[4 91 92] PGH

2





2

and H2







[104ndash113][81 108 114ndash121]

Cancer MDA4-HNE



[72 79 109 123 135 137ndash141][72 104 109 131 135 138 139 142ndash144]

Diabetes MDA4-HNE

[79 109 123 140 145ndash150][131 135 142 143 151ndash156]




[81 108 114ndash121][72 114 131 135 142 170ndash174]

PUFAAA



Monocyclicperoxide

HHTMDA

Malonicsemialdehyde




1

2

3 33

4

546

7

Cyclization

CO2 + H2O

O2

O2

+ H+

H+

H+


Radical∙

PUFA-radical∙

2O2

PGG2

PGH2

TXA2


2

(TXA2












LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















2


2


2

)[4 91 92] PGH

2





2

and H2







[104ndash113][81 108 114ndash121]

Cancer MDA4-HNE



[72 79 109 123 135 137ndash141][72 104 109 131 135 138 139 142ndash144]

Diabetes MDA4-HNE

[79 109 123 140 145ndash150][131 135 142 143 151ndash156]




[81 108 114ndash121][72 114 131 135 142 170ndash174]

PUFAAA



Monocyclicperoxide

HHTMDA

Malonicsemialdehyde




1

2

3 33

4

546

7

Cyclization

CO2 + H2O

O2

O2

+ H+

H+

H+


Radical∙

PUFA-radical∙

2O2

PGG2

PGH2

TXA2


2

(TXA2












LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















[104ndash113][81 108 114ndash121]

Cancer MDA4-HNE



[72 79 109 123 135 137ndash141][72 104 109 131 135 138 139 142ndash144]

Diabetes MDA4-HNE

[79 109 123 140 145ndash150][131 135 142 143 151ndash156]




[81 108 114ndash121][72 114 131 135 142 170ndash174]

PUFAAA



Monocyclicperoxide

HHTMDA

Malonicsemialdehyde




1

2

3 33

4

546

7

Cyclization

CO2 + H2O

O2

O2

+ H+

H+

H+


Radical∙

PUFA-radical∙

2O2

PGG2

PGH2

TXA2


2

(TXA2












LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


























LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




















LA

9-HPODE

15-LOX

Alkenal derived

4-HNE

HP-Lyase Alkenal OX

4-HPNE

Peroxygenase

GS-HNE

ALD

H

DHNHNA

GH-HNA

ALD

H

ADH

ADHGSH

GH-DHN

CYP

9-OH-HNA





PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















PUFAlipoic acid

9 10 dioxetane

4-HPNE


Fragmentation

Reduction

4-HPNE

Peroxy dioxetane

4-HNE 4-HNE

Peroxycyclization

Fragmentation

Rearrangement

21 3

54

H+

H+H+

H+


13-Lipid radical∙



O2

O2O2

O2

O2

4-HNE 4-HNE

9-Lipid radical∙




4-HNE

120573-Scission








to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















to stress


Physiologicallevels



4-HNE ismetabolized



states



death

Lowlevels

Mediumlevels

Highlevels

Very highlevels


4-HNE 4-HNE 4-HNE

4-HNE 4-HNE 4-HNE



Cell die


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



























2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















2

O2
















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


























50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






















50









2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






















2





+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















+

LH

LH

Cumoxyl

Cumoperoxyl


3

1

5

4

2

1

3

H3C C

OH

CH3H3C C CH3H3C C

OOH

CH3

H3C C

OH

CH3 H3C C CH3 H3C C

OOH

CH3

OO∙

O∙




L∙

L∙

O2

M(n+1)Mn

minusOH















5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


























5 Conclusions




Acknowledgments


References







































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















































































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of













































































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of












































































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of










































































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of











































































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of










































































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of










































































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of







































































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of







































2











































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















































































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of















































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Review Article Lipid Peroxidation: Production, Metabolism ...downloads.hindawi.com/journals/omcl/2014/360438.pdf · Review Article Lipid Peroxidation: Production, Metabolism, and

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