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
M . Pharm (Pharmacology)
HCC is the most common type of liver malignancy and 4th most common cause of
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 :
Hepatitis viral infection
Excessive alcohol consumption and smoking
Exposure with chemical carcinogens
Air and water pollutants
Bone marrow depression
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
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
popularly known as “Chiranji” in Hindi
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 3540ºC 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
Grouping of the selected animals
Experimental animals are randomly divided into 5 groups
GROUP - І
GROUP – ІІ (DEN
GROUP - ІІІ
(DEN+TEST 1 ;
GROUP – ІV
(DEN+TEST 2 ;
GROUP - V
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;
After 21 days of observation the overnight fasted animals were
sacrificed by chemically induced euthanasia using chloroform.
Blood samples were collected by
serum was obtained from the blood by
centrifugation at 4,000 rpm for 20
minutes. The supernatant is used for
Serum enzymes like
SGPT, SGOT, ALP, total protein
and bilirubin values were
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.
Result and graphs:
Table no. 1. Data showing preliminary Phytochemical screening of the ethanolic extract of
Buchanania lanzan leaves extract.
Ethanolic extract of
Buchanania lanzan leaves
Test for Alkaloids
Test for Steroids
Test for Tannins
Test for Saponin
Test for Gallic acid
Test for Flavonoids
Test for Carbohydrates
Test for Gum and Mucilage’s
Test for Glycoside
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)
SGOT & SGPT level (IU/L)
Effect of Buchanania lanzan on SGOT and SGPT level in DEN induced Hepatocarcinoma in rats
* Reitman et.al 1957
Effect of Buchanania Lanzan on Biochemical parameters in DEN
induced Hepatocarcinoma in rats.
ALP and TP level
Effect of Buchanania lanzan on bilirubin level in DEN induced Hepatocarcinoma
Bilirubin level (mg/dl)
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
Group III (p<0.01) vs. Group II
Group IV (p<0.01) vs. Group II
Group V (p<0.01) vs. Group II, N=6
SOD = µ moles of MDA/min/mg protein
CAT = µ moles of H2O2 consumed/min/mg protein
LPO = µ moles of MDA/min/mg protein
Effect of Buchanania Lanzan on SOD and CAT level in DEN induced Hepatocarcinoma in
SOD and CAT level
Effect of Buchanania Lanzan on LPO level in DEN induced Hepatocarcinoma in rats.
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.
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
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.