buchanania lanzan hepatocarcinoma presentation

Abstract Reference Number : OPP 030

Effect of total extract of Buchanania lanzan leaves against Hepatocellular

Carcinoma(HCC) in Diethyl nitrosamine induced tumor model

Sumithra M*, Anbu J, Srota Dawn, Ashwini Anjana, Nazeer Ahmed,

Ravichandiran V.

Department of Pharmacology, Vels University

Pallavaram, Chennai-600117

Presenting Author

Srota Dawn

M . Pharm (Pharmacology)

Introduction:

HCC is the most common type of liver malignancy and 4th

most common cause of cancer mortality.

Worldwide incidence >600,000 cases per year

Liver cancer is the most rapidly increasing cancer in the U.S.,

Asia and Africa.

19,160 new cases and 16,780 deaths in 2007

More common in men than women (4:1)

For resection, rate of recurrence can be as high as 50% at 2

years

Only 12% are eligible for resection or for transplant

80%-90% of HCC cases occur in cirrhotic livers

Factors causing HCC :

Environmental factors

Hepatitis viral infection

Food additives

Excessive alcohol consumption and smoking

Alfatoxins

Exposure with chemical carcinogens

Air and water pollutants

Management of HCC:

Liver transplantation

Resection

Tumour ablation

Radiofrequency thermal ablation

Alcohol injection

Chemoembolization

Targeted molecular therapy

Chemotherapy

Regional

Systemic

Bone marrow depression Lymphocytopenia, Stomatitis

Gastro intestinal tract disorders like Diarrhoea,

Nausea, Vomiting

Skin problems like Alopecia, Dermatitis

Oligozoospermia, Impotency in males

Abortion and Fetal damage in females

Generation of Secondary cancers specially leukemia,

lymphoma

TOXIC EFFECTS OF ANTICANCER AGENTS

Aim :

Here the anticancer potential of ethanolic extract of

Buchanania lanzan (Anacardiaceae family) leaves was

evaluated against Diethyl nitrosamine (DEN) induced

hepatocarcinoma in male wistar rats.

The aim of this study is to reduce the toxicity and adverse

effect of cytotoxic drugs by the use of drugs from

phytochemical sources.

About the Plant:

Buchanania lanzan (Anacardiaceae family) is an evergreen deciduous tree, growing 50 ft tall. popularly known as “Chiranji” in Hindi

Uses:

Buchanania lanzan has folklore evidence for its effective role in curing asthma, cough, skin diseases, antioxidants and antitumor effect. All parts of the plant are used for the treatment of various diseases. The oils extracted from the seed are used for reduction of granular swelling of neck The kernel is used to treat skin diseases like itching and as prickly heat also The gum obtained from the bark of Buchanania lanzan used for treating diarrhoea and intercostals pains. Leaves of Buchanania lanzan tree are used for promoting wound healing

Work plan and procedure

Collection and authentication of plant material

Extraction procedure (soxhlet extractor, 70% ethanol, temperature 35-40ºC for 72 hrs.); preliminary phytochemical

studies

Selection of animals(30 adult male wistar rats, 100-150 g); acclimatized for 2 weeks, 12 hrs. day/night cycle, pallet food,

water ad libitum ;

experimental procedure and protocol used in this study was reviewed and approved by Institutional Animal Ethics

Committee (IAEC)

The doses are fixed after performing the acute toxicity studies according to the OECD guideline 423

Continued….

Experimental animals are randomly divided into 5 groups (n=6)

GROUP - І (control)

GROUP – ІІ (DEN

alone)

GROUP - ІІІ

(DEN+TEST 1 ; 200 mg/kg)

GROUP – ІV

(DEN+TEST 2 ;

400mg/kg)

GROUP - V (DEN+ 5

Flurouracil)

Grouping of the selected animals

DEN (in DMSO) is given intraperitonially at a dose of 50 mg/kg once in a week for a period of three weeks to GROUP – ІІ,

GROUP – ІІІ, GROUP - ІV and GROUP - V ; The test drug is given for 15 days;

Continued.........

After 21 days of observation the overnight fasted animals were sacrificed by chemically induced

euthanasia using chloroform.Blood samples were collected by cardiac

puncture

serum was obtained from the

blood by centrifugation at 4,000 rpm for 20

minutes. The supernatant is

used for estimation of

Serum enzymes like SGPT, SGOT,

ALP, total protein and

bilirubin values were evaluated.

determination of antioxidant enzymes, liver was dissected

Using ice cold saline and

homogenate was prepared (0.1M

Tris–HCL buffer, pH 7.4) , centrifuged

and the supernatant was

used for the estimation of

Tissue LPO , SOD and CAT

level were estimated.

All data obtained from the experiment were represented as mean ± SEM and analyzed by One way ANOVA and for multiple comparisons of groups with post hoc Bonferroni’s test was used.

Sl. No. Phytochemical testEthanolic extract of Buchanania lanzan

leaves extract

1. Test for Alkaloids +ve

2. Test for Steroids -ve

3. Test for Tannins -ve

4. Test for Saponin -ve

5. Test for Gallic acid +ve

6. Test for Flavonoids +ve

7. Test for Carbohydrates -ve

8. Test for Gum and Mucilage’s -ve

9. Test for Glycoside +ve

Result and graphs:Table no. 1. Data showing preliminary Phytochemical screening of the ethanolic extract of Buchanania lanzan leaves extract.

Sl. no.

Groups (n=6)

SGOT (IUL-1) SGPT (IUL-1) ALP (IUL-1)TP

(mg/100g) Bilirubin

(mg/dL)

1. Normal control

51.47±0.33

98.25±0.27 220.25±0.25 767.90±0.230.460±0.0

22

2. DEN alone 103.13±0.45 255.51±0.15 295.33±0.62 832.16±0.52 1.517±0.093

3.DEN+200mg/kg extract(T1)

91.21±0.23 242.98±0.34 288.12±0.33 818.32±0.43 0.510±0.06

4.DEN+400mg extract(T2)

84.45±0.65 180.43±0.33 252.65±0.22 792.21±0.21 0.536±0.12

5. DEN+ 5FU 81.11±0.54 167.55±0.56 247.55±0.93 788.34±0.99 0.510±0.05

Table no. 2. Effect of ethanolic extract of Buchanania lanzan leaves on Biochemical parameters in DEN induced Hepatocarcinoma in rats.

All values are presented as mean ± SEM. Group II was compared with group I and all values were significant (p< 0.001); group III, group IV and group V were compared with group II and all values were significant. (p< 0.001). P values were calculated by one way ANOVA analysis with post hoc Bonferroni’s test. (N=6)

Normal control DEN alone DEN+200 mg/kg extract(T1)

DEN+400 mg extract(T2)

DEN+ 5FU0

50

100

150

200

250

300

51.47

103.13 91.21 84.45 81.1198.25

255.51 242.98

180.43 167.55

Effect of Buchanania lanzan on SGOT and SGPT level in DEN induced Hepa-tocarcinoma in rats

SGOT (IUL-1) SGPT (IUL-1)

Groups

SG

OT

& S

GP

T l

eve

l (I

U/L

)

Graphical representation

* Reitman et.al 1957

Normal control DEN alone DEN+200 mg/kg extract(T1)

DEN+400 mg/kg extract(T2)

DEN+ 5FU0

100

200

300

400

500

600

700

800

900

220.25295.33 288.12 252.62 247.55

767.9832.16 818.32 792.21 788.34

Effect of Buchanania Lanzan on Biochemical parameters in DEN induced Hepatocarcinoma

in rats.

ALP (IUL-1)TP (mg/100g)

Groups

ALP

an

d T

P l

eve

l

Normal control DEN alone DEN+200mg/kg extract(T1)

DEN+400mg/kg extract(T2)

DEN+ 5FU0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

0.46

1.517

0.51 0.536 0.51

Bil

iru

bin

le

vel

(mg

/dl)

Groups

Effect of Buchanania lanzan on bilirubin level in DEN induced Hepatocarcinoma in rats.

Sl. no.

Groups(n=6)

SOD CAT LPO

1. Normal control 8.89±0.12 63.21±1.45 0.095±.003

2. DEN alone 4.44±0.10 41.85±1.33 0.219±0.004

3.DEN+200mg extract(T1)

5.11±0.21** 48.55±1.09** 0.115±0.013

4.DEN+400mg extract(T2)

7.65±0.33 56.09±0.99 0.178±0.02**

5. DEN+ 5FU 7.99±0.55 57.99±1.93 0.099±0.003

Table no. 3. Effect of Buchanania Lanzan on Antioxidant level in DEN induced Hepatocarcinoma in rats.

All values are presented as mean ± SEM. Group II was compared with group I and all values were significant (p<0.001)

Group III (p<0.01) vs. Group IIGroup IV (p<0.01) vs. Group IIGroup V (p<0.01) vs. Group II, N=6SOD = µ moles of MDA/min/mg proteinCAT = µ moles of H2O2 consumed/min/mg proteinLPO = µ moles of MDA/min/mg protein

Normal control DEN alone DEN+200mg/kg extract(T1)

DEN+400mg/kg extract(T2)

DEN+ 5FU0

10

20

30

40

50

60

70

8.89

4.44 5.117.65 7.99

63.21

41.8548.55

56.09 57.99

SOD CAT

Groups

SO

D a

nd

CAT level

Effect of Buchanania Lanzan on SOD and CAT level in DEN induced Hepatocarcinoma in rats.

Normal control

DEN alone DEN+200mg/kg extract(T1)

DEN+400mg/kg

extract(T2)

DEN+ 5FU0

0.05

0.1

0.15

0.2

0.25

0.095

0.219

0.115

0.178

0.0990000000000001

Groups

LP

O l

eve

l

Effect of Buchanania Lanzan on LPO level in DEN induced Hepatocarcinoma in rats.

Discussion:

Elevation of the plasma levels of cytoplasmic and

mitochondrial enzymes are sensitive indicators of liver damage.

Elevation of serum SGPT, SGOT, ALP, TP, LPO and bilirubin is known

effect of DEN toxicity which specially affects the liver and activities

of ALP, SGPT, SGOT, and LPO are most commonly used biochemical

markers of liver damage. Lipid peroxidation plays an important role

in carcinogenesis

There is a significant dose dependent reduction in SGOT ,

SGPT, ALP, TP level and SOD, CAT levels are increased in dose

dependent manner in the B. lanzan treated groups.

The low dose of extract shows a better effect in reduction of

bilirubin and LPO level than the high dose.

Conclusion:

The preliminary phytochemical studies reveal the presence of

flavonoids, Alkaloids, Gallic acid and Glycoside in total extract of

Buchanania lanzan.

After studying many journals we can say that Flavonoid and alkaloids

has a chemo preventive role in cancer. Therefore the possible

mechanism of anticancer of Buchanania lanzan may be due to

flavonoid or alkaloid content and further study has to be carried out

to study the exact mechanism behind it and isolation of the active

constituents present in the extract.

Acknowledgments

The authors are thankful to Vels University, Pallavaram, Chennai –

600117, India,

for providing the necessary laboratory facilities for this study.

I want to thank my teacher Sumithra M. for giving me the

opportunity to present this paper and I also want to thank the

authority of NATIONAL CONFERENCE ON HERBAL MEDICINE

(NCHM’13) for encouraging the students and research scholars

and for giving a platform to present their views.

THANK YOU