HEPATOPROTECTIVE & SYNERGISTIC

EFFICACY OF POLYHERBAL
FORMULATION FROM TINOSPORA
CORDIFOLIA AND MURRAYA KOENIGII L
AGAINST CARBON TETRACHLORIDEINDUCED (CCL4) TOXICITY: IN VIVO STUDY
V.NISHEETHA (12FJ1S0115)
UNDER THE GUIDANCE OF G.KIRANMAI
(ASSISTANT PROFESSOR OF
PHARMACOLGY)
SSJ COLLEGE OF PHARMACY.

INTRODUCTION
Liver is a vital organ has a wide range of
functions in the body including carbohydrate,
protein and fat metabolism, detoxification,
secretion of bile and storage of vitamins,
synthesis of plasma protein and glycogen
storage .
The liver is also prone to many diseases mostly
included are infectious such as hepatitis A,B,C,E,
alcohol damage, fatty liver, cirrhosis, cancer,
drug damage especially acetaminophen, cancer
drugs .

Carbon tetrachloride (CCl4) induced hepatotoxicity model

is widely used model for the study of hepatoprotective
effects of drugs and plant extracts .
Natural remedies from medicinal plants are considered to
be effective and safe alternative treatment for liver
diseases. Therefore, many folk remedies from plant origin
are tested for its potential antioxidant and
hepatoprotective liver damage in experimental animal
model.
Polyherbal formulation from Tinospora cordifolia and
Murraya koenigii L against Carbon TetrachlorideInduced (ccl4) toxicity, is studied .

HERBAL PRODUCTS
TINOSPORA CORDIFOLIA
CLASSIFICATION
KINGDOM- PLANTAE
DIVISION: MANGOLIOPHYTA,
CLASS: MAGNOLIOPSIDA,
ORDER: RANUNCULALES,
FAMILY: MENISPERMACEAE,
GENUS: TINOSPORA,
SPECIES: T. CORDIFOLIA,
BOTANICAL NAME: TINOSPORA CORDIFOLIA

MURRAYA KOENIGII
CLASSIFICATION
KINGDOM: PLANTAE
DIVISION: ROSIDS
CLASS: EUDICOTS
ORDER: SAPINDALES
FAMILY: RUTACEAE
GENUS: MURRAYA
SPECIES: MURRAYA KOENIGII
BOTANICAL NAME: MURRAYA KOENIGII

AIMS AND OBJECTIVES

Preparation of ethanolic extract from Tinospora cordifolia

and Muraya konigii and their phytochemical analysis

To study antioxidant activity of ethanolic extract of

T.cordifolia and M.Koenigii

Induction of hepatotoxicity by CCl4
Evaluation of hepatoprotecive efficacy of the plant extracts

in rats by biochemical analysis.

Pathophysiological studies of various organs and organ

weights.

MATERIALS AND METHODS

1.Preparation of plant extracts
2. Preliminary phytochemical analysis.
3.Antimicrobial activities.
4. Antioxidant activity
5. Animals and experimental design
6. Experimental Design
7. Carbon tetrachloride induced hepatotoxicity

8. Biochemical and histopathological studies

9. Assesment of lipid peroxidation.
10. Serum ALT, AST and Total Bilirubin levels.
11. Histopathology .

Preparation of plant extracts

The leaves of Tinospora cordifolia and

Muraya Koenigii (MK) were collected

from local market.
Leaves were shade dried and pulverized to a

coarse powder and extracted with ethanol. The

filtrate obtained was evaporated to dryness at
50-65C in a rotary vacuum evaporator to
obtain a dark colored molten mass.

Phytochemical analysis :
Screening is done to find the presence of the
active ingredients in the leaf extracts with
different solvents.
Antimicrobial activity :
Antimicrobial activity indicates that biological activity of
crude extracts, In that we are both Gram negative, Gram
positive bacteria for determination of MIC (Minimum
Inhibitory Concentration), MBC and antifungal activity of
the extracts will be estimated.

Animals design:
SEX- Male SD rats
WEIGHT 180 200 gm.
NO.OF GROUPS 6 groups ( in each group 4
animals.)
TOTAL NUMBER OF ANIMALS 24 male sd rats.

Experimental design.
Group 1: Normal control
Group 2: CCl4 control group
Group 3: Standard drug sylimarin 30mg/kg body

weight+ CCl4
Group-4 : TC-150mg/kg + CCl4
Group-5: MMK 150mg/kg + CCl4
Group 6: 300 mg herbal formulation/kg body

weight+ccl4

PROCEDURE
Animal groups were divided and treated

accordingly.
1st group Treated normally with orally 0.5% of
CMC for seven days, and then intraperitonially
injected with 10 ml / kg body weight olive oil.
2nd group - served as CCl4 hepatotoxicity control

and was orally given 0.5 % CMC for seven days

and then intrateritoneally intoxicated with ccl4
10ml/kg body weight (0.1% in olive oil).

Third group served as standard group and

received reference drug, silymarian (25 mg / kg
/day, p.o) for seven days prior to CCl4
intoxication.
Fourth group of animals were pretreated with 150
mg / kg body weight of plant extract during the
seven days, and then intoxicated with CCl4
10ml/kg body weight (0.1% in olive oil).

All animals were sacrificed under mild ether anesthesia

16 hours after ccl4 injection, and serum was separated at
2500 rpm for 15 min and biochemical investigations
were carried out.
Liver was dissected out and used for biochemical
determinations and liver tissue slices were collected for
histopathological studies and blood samples were
collected immediately.

OBSERVATION AND ANALYSIS OF THE STUDY.

Biochemical parameters like serum glutamate
pyruvate transaminase (SGPT), serum glutamate
oxaloacetate transaminase (SGOT), total proteins,
albumin and serum bilirubin ( SB ) were assayed.
The antilipidperoxidant effect of two different

extracts and combination of herbal formulation

was studied in vitro.

Serum ALT, AST and Total Bilirubin levels

There was a significant increase in serum enzymes ALT
and AST activity of CCl4 treated groups when
compared with that of normal control.
Histopathology
Animals were sacrificed to remove liver. The liver was
fixed in Bouins solution for12 hr, and then embedded in
paraffin using conventional methods (Galighor 1976) ,
cut into 3m thick sections and stained using
haematoxylinbosin dye. The sections were then
observed for histopathological changes.

Conclusion:
Hence the comparison is made between the

groups of animals divided and observed for the

serum levels and histopatology results and animal
is sacrificed.

References
"Murraya koenigii information from NPGS/GRIN".
www.ars-grin.gov. Retrieved 2008-03-11. Jump up ^
Arulselvan P, Senthilkumar GP, Sathish Kumar D,
Subramanian S (Oct 2006). "Anti-diabetic effect of
Murraya koenigii leaves on streptozotocin induced
diabetic rats". Pharmazie 61 (10): 8747. PMID
17069429.
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