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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
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