Anti Oksidan vs Free Radicals

Free Radicalsvs Anti Oksidan

Noor WijayahadiThe Price for Living: Free Radical Generation and Defense Mechanism

Bacteria Anti-biotic

Free Radicals Anti-oxidants(Oxidants)

RECORDS IN LIFE SPAN

Kura-kura 170 tahunKucing30 tahunManusia116 tahunAnjing27 tahunKerang80 tahunSapi20 tahunKakatua70 tahunKelinci18 tahunGajah70 tahunAyam14 tahunBurung hantu68 tahunTikus5 tahunKuda62 tahunMencit5 tahunChimpanse50 tahunCoro1 tahunGorilla48 tahunNyamuk5 bulanBeruang47 tahunLalat70 hariMerpati30 tahun

THEORIES OF AGING PROCESS :Genetic Clock TheorySomatic Mutation TheoryRate of Living or Metabolism TheoryImmune System Destruction Theory Free Radicals TheoryLain-Lain:kesepian, banyak pacar, banyak hutang/kredit, banyak pasien, dll

What are free radicals?Adalah molekul yang kelebihan elektron sangat reaktif

Reactive Oxygen Species (ROS) Reactive Nitrogen Species (RNS) - NOReactive Metabolites or Intermediates produk metabolisme obat, toxin, pollutant, merokok, dll

Origins of reactive oxygen, nitrogen and chlorinating species

Functions of oxidant

1. Reactive Oxygen Species (ROS)Superoxide (O2.-)Hydrogen Peroxide (H2O2)Hydroxyl Radical (OH.) - product of Fenton reaction catalyzed by free Fe and CuSinglet Oxygen (1DgO2) - oxygen at an excited state, requiring photosensitizers and photons

Sumber oxygen free radicalsIn mitochondria:- generation of energy - ATP- glucose, fatty acids, amino acids- O2 2H2O 4e-- leakage of O2-. (superoxide) H2O2 (hydrogen peroxide)

In Smooth Endoplasmic Reticulum (microsome)- detoxification (cytochrome P-450s)- toxins, drugs and xenobiotics- O2 + RH R-OH and H2O- leakage of O2-. - metabolic activation - X.

In Smooth Endoplasmic Reticulum (microsome)- eicosanoids synthesis - substrate: arachidonic acid (AA)- O2 + AAs prostaglandins, thromboxanes and leukotrienes- leakage of O2-.

In Peroxisomes- containing oxidases for degradation of various substrates- glucose, amino acids, xanthine, etc.- requires O2- byproduct is H2O2

In Cytoplasm- nitric oxide (NO.) production from Arginine- functions as a biological messenger- in brain, vascular endothelial cells, and macrophages- NO. + O2-. ONOO. (peroxynitrite)

2. NO: a Biological MessengerNO is a neurotransmitter (brain- bNOS)NO regulates blood pressure (vascular endothelial cells- eNOS)NO is a cytotoxic agent (macrophages- iNOS)

13413.1. Sources of Superoxide Anion Enzymic reactions NADH oxidase NADPH-P450 reductase xanthine oxidase

Cellular sources leukocytes and macrophages mitochondrial electron transfer microsomal monooxygenase Environmental factors ultraviolet light X-rays toxic chemicals aromatic hydroxylamines aromatic nitro compounds insecticides chemotherapeutic agents

1342

13433.3. Sources of Hydroxyl Radical Most of the HO. generated in vivooriginates from thebreakdown of H2O2 via a Fenton reaction

13433.3. Sources of Nitric Oxide Nitric oxide is generated by a Ca-dependentmixed function oxidase nitric oxide synthase (NOS)in mammalian tissuesNOS

13433.3. Sources of Nitric Oxide Nitric Oxide Synthases (NOS) in mammalian tissues

Types of NOSNOS ICentral and peripheral neuronal cellsCa+2 dependent, used for neuronal communicationNOS IIMost nucleated cells, particularly macrophagesIndependent of intracellular Ca+2Inducible in presence of inflammatory cytokinesNOS IIIVascular endothelial cellsCa+2 dependentVascular regulation

13433.3. Sources of Nitric Oxide Nitric Oxide Synthases (NOS) in mammalian tissuesMayor Domainsoxygenase domainreductase domaincalmodulin

Oxygen MetabolismMitochondria e - transportH2O2H2OO2O2.-SERP-450PHSAAPGsLTsH2OO2O2.-PeroxisomeOxidasesO2MembraneMPxNOSNO.H2O2O2.-NO.HOCl-PSO21O2hn

Oxygen Consumption and Oxidative Stress O2 Consumption: 4 x 1010 O2/cell/hr

H2O2 Production: 3.24 x 109 H2O2/cell/hr

Mitochondria - 13.6%Microsomes - 47.7%Peroxisomes - 34.1%Cytosol - 4.6%

Boverls, et al. Biochem. J. 128:617 (1972) where rat liver at 22 0C was used

Oxidative DamageFree Radicals

Proteins Lipids DNA/RNA (-SH)(R-OO.) (-OH.)

Severity of Oxidative Stress & Biological ConsequencesSeverity of Oxidative StressBiological ConsequencesA. Low level & gradualAgingB. Medium level & rapidCarcinogenesis MutagenesisC. Large level & rapidDeath, Stroke, Trauma, Ionizing irradiation

Free Radical Involvementin Pathophysiological ConditionsAdriamycin cardiotoxicityAIDSAdult Resp Distress SyndAgingAlcoholismAlzheimers DiseaseAmyotrophic Lateral SclerosisAtherosclerosisDiabetesCancerExerciseFavismIron OverloadMyocardial InfarctionOxygen ToxicityParkinsons DiseaseRadiation TherapySmokingStrokeTrauma

Breathing is bad for youAll breathing generates oxygen radicals, which are the main sources of mutation in DNA, leading to cancer, birth defects, and various peculiarly shaped molecules in urine. Breathing has been observed three minutes before death in 100% of fatalities. We urge everyone to stop breathing until further research has been carried out.

Oxidative StressANTIOXIDANTSOXIDANTSOxygen Free RadicalsNitrogen Free RadicalsSpecific EnzymesVitamin E, Vitamin C,Carotenoids, Selenium

Free Radical Defense SystemAntioxidant Enzymes Antioxidant QuenchersAntioxidant from Foods nutrients/non-nutrients preventative mechanisms repair mechanisms physical defences antioxidant defences

AntioxidantsPrevents the transfer of electron from O2 to organic moleculesStabilizes free radicalsTerminates free radical reactions

Antioxydant defence mechanisms

1346Primary Antioxidant DefensesSpecific Antioxidant EnzymesSuperoxide dismutaseCatalaseGlutathione PeroxidasesGlutathione TransferasesSmall Antioxidant MoleculesVitamin EVitamin CCoenzyme QUric AcidCarotenoids

Antioxydant network to maintain redox balance

O2H2O2H2O + O2Mn SODCatalaseGSH PeroxidaseCuZnSODOHFe2+Antioxidant Enzymes

Antioxidant Enzymes-1Superoxide Dismutase (SOD) to get rid of superoxide produced from electron transport chain, the product is hydrogen peroxide.MnSOD (mitochondria).CuZn SOD (cytosol).

Antioxidant Enzymes - 2Glutathione Peroxidase (GSH PX) to get rid of hydrogen peroxide (H2O2) and some lipid peroxide. It requires reduced glutathione (GSH) as substrate and produces oxidized glutathione (GSSG) as product. A cytosolic enzyme.Catalase to get rid of hydrogen peroxide produced in peroxisome. Antioxidant Enzymes - 3

Glutathione Synthesis ProteinMethionineCysteineGlutamateg-GlutamylcysteineGlycine

Supplementation of Cysteine Prodrugs ProteinMethionineCysteineSupplementation:OTCNACMETGSH Glutamateg-GlutamylcysteineGlycine

Cysteine ProdrugsNAC N-Acetyl-CysteineOTC - L-2-oxothiazolidine- 4- carboxylateGSH GlutathioneMET Methionine analogsothers

Conversion of OTC to Cysteine by OxoprolinaseSNHHCOO-OCOO- SCH2CHCOO-NH3ADP + PiHSCH2CHCOO-NH3L- CysteineL-2-oxothiazolidine- 4- carboxylate (OTC)Carboxy- L- cysteineOxoprolinaseATPCO2

Antioxidant QuenchersCellular proteins which chelate pro-oxidant minerals (iron and copper or others)Transferrin iron transport proteinFerritin iron storage proteinMetallothionein minerals and heavy metals (Zn/Cu/Cd/Hg)Ceruloplasmin copper transport and storage

Antioxidants From FoodAntioxidant nutrients vitamin Evitamin C(vitamin A?)beta-carotenePhytochemicals antioxidants from plants

NutrientsNon-nutrientsEnergy, buildingmaterialsFactors regulating metabolismLipidProteinCarbo-hydrateVitaminsMineralsEFA & non-EFAGlucoseWaterPhyto- chemicalspigmentsAntioxidants

Fibers

Other food components

FoodGSHCysFunctional food or Neutraceuticals

C, E, b-CarZn, SeEAA & non-EAA

Discovery of Other Functions of PhytochemicalsAnti-oxidantAnti-inflammatoryAnti-estrogenicAnti-allergic

Anti-cholesterolemicAnti-hemorrhagicAnti-mutagenicAnti-neoplastic

Antioxidant NetworkVitamin E

Vitamin E radicalsVitamin C

Vitamin C RadicalsGSH

GSSGPhytochemicals

Phytochemicals-O

Oxidative DamageOxidative Stress NF-kB ActivationGene Transcription Cytokines, Chemotactic FactorsInflammationROS Effect of Oxidative Stress on Oxidative Stress Gene Expression (NFkB Activation)

Oxidative DamageOxidative Stress NF-kB ActivationGene Transcription Cytokines, Chemotactic FactorsInflammationROS Antioxidantsvitamin EEffect of antioxidants on the inhibition of NFkB activation

Prevention of Free Radical-Mediated Chronic DiseasesCell DeathGeneticEnvironmentalViralDietaryROS DevelopmentalHormonalNF-kB ActivationGene Transcription Cytokines, Chemotactic FactorsInflammationROS AntioxidantsAnti-InflammatoryAmplification

PHAGOCYTE

HEALTHY PROSTATE TISSUEInflammation is a source of endogenous mutagenic chemicals

BIOMARKER KERUSAKAN OKSIDATIF

Hidup = Keseimbangan Pembentukan Radikal-Bebas vs Mekanisme Pertahanan

**Origins of reactive oxygen, nitrogen and chlorinating species************Protein provides the supply of cysteine in the GSH synthesis. Cysteine is also the limiting amino acid in the synthesis. So when dietary protein level or the body protein status is low, cysteine supply would be limited and GSH synthesis would be reduced. *Therefore, in order to increase glutathione concentrations without feeding high protein diet during the early PM rehabilitation, one needs to increase cysteine supply by supplementing cysteine prodrugs. Commonly used cysteine prodrugs include NAC (N-acetylcysteine), OTC (L-2-oxo-4-thiazolidine-carboxylate ), methionine and glutathione itself. Supplementation of cysteine itself is not used because cysteine is chemically unstable and also can be toxic at high doses.

***