INTRODUCTION:
The liver is a
vital organ of paramount importance involved in the maintenance of metabolic
functions and detoxification from the exogenous and endogenous challenges like xenobiotics, drugs, viral infections and chronic
alcoholism. Hepatic damage is associated with distortion of these metabolic
functions; the liver damage is always associated with cellular necrosis,
increase in tissue lipid per oxidation and depletion of tissue GSH levels. In
addition serum levels of many biochemical markers like SGOT, SGPT,
triglycerides, cholesterol, Bilirubin and protein are
elevated1. Liver disease is still a worldwide serious health
problem. In spite of phenomenal growth of modern medicine, there are no
synthetic drugs available for the treatment of hepatic disorders. Nature has
provided an excellent storehouse of remedies to cure all the ailments of
mankind. Modern research is now focusing greater attention on the generation of
scientific validation of herbal drugs based on their folklore claim. In this
modern era, a large Indian population still relies on the traditional system of
medicine, which is mostly plant based1,2.
Many plants
conveniently available in India are used in traditional folklore medicine for
the treatment of hepatic diseases. Among them Butea monosperma lam (Fabeceae)
also known as flame of forest is wild, medium sized tree found throughout the
deciduous forests and open areas was claimed to possess hepatoprotective
property3,4. In the literature B.
monosperma is ascribed to have many medicinal
properties. Its flowers are used in the
treatment of hepatic disorders and viral hepatitis, diarrhoea
and anti implantation activity5. The roots are used to night
blindness, helminthiasis, piles, ulcers and tumours. The bark is reported to antitumour
and anti ulcer activities, the root bark is used as an aphrodisiac, analgesic
and anthelmentis, the leaves possess antimicrobial
property6. It was found that these plants are found to posses polyphenolic constituents like flavonoids.
The B. Monosperma
barks contain Kino- tannic acid, Gallic
acid, pyrocatechin, butrin,
palasitrin, alanind,
allophonic acid, butolic acid, cynidin,
histidine, lupenone, lupeol, miroestrol, palasimide and shelloic acid 7.
Flavonoids are reported to have anti-inflammatory, antihepatotoxicity and antiulcer actions. They are potent
antioxidants and have free radical scavenging abilities 8. Hence the
present study was carried out to determine effect of methanolic extract of B.monosperma lam
stem bark on Carban tetrachloride induced hepatotoxicity and Brewer’s yeast induced pyrexia in rat
models.
MATERIALS AND METHODS:
Plant Material and Extraction
The Stem Bark of B.monosperma (lam) was collected from the areas around Madurai, Tamilnadu, India in the month of August 2010. The plant was identified and authenticated by
Dr. S. Baburaj, Botanist, Thyagarajar
College, Madurai. The stem barks were
dried under shade for 30 days and homogenized to get coarse powder and was
stored in an air tight container. 1kg of dried stem bark powder of B.monosperma was
extracted with 80% of methanol (65˚-75˚C) in a soxhlet
apparatus for 24 h. The methanolic extract was concentrated under reduced
pressure in rotary evaporator 9. The methanolic extract of B.monosperma
(MEBM) obtained was reddish brown in colour and the
yield was found to be 5% w/w. For pharmacological studies the dried extract was
suspended in Normal saline to make the required dose.
Animals
Male Albino rats of Wister strain (150-200gm) were used
and were housed in polypropylene cages at room temperature (25
2oC) proper humidity (44-55%) conditions and
maintained on 12h day-night cycle. The animals were fed with commercial rat
pellets (Amrut laboratory animal feed Ltd. Bangalore)
and were given water ad libitum. The animals were acclimatized for a period of
20 days prior to performing the experiment. The experimental protocol and all
the procedures were approved by Institutional Animal Ethical Committee of Ultra
College of Pharmacy (UCP/IAEC/2010/52).
Hepatoprotective
activity
Carbon
tetrachloride induced hepatotoxicity
The rats were divided into five groups of 3 animals
each. Group I served as normal control and received normal saline
(2ml/kg, p.o); Group II was administered with CCl4
in liquid paraffin 30% v/v (1ml/kg, i.p). Group III
was treated with the standard Liv-52 (1ml/kg, p.o).
Group IVandV was treated with MEBM 200 and 400 mg/kg,
p.o respectively 10. Group III, IV and V
received CCl4 in liquid paraffin 30% v/v (1ml/kg, i.p) for every 72 hours. The respective drug treatments
were carried out for a period of 10 days 11, 12. On 11th day the blood
samples were withdrawn from all animals under mild ether anaesthesia
through the retro-orbital puncture. The blood Samples were allowed to clot for
10min at room temperature and then centrifuged at 2500rpm for 20min at 30°C and
serum was stored at 4-80C for the estimation of various biochemical
parameters. Biochemical
parameters such as SGOT, SGPT, ALP, total bilirubin
and total protein were analyzed according to the standard procedures13-16.
Antipyretic activity
Antipyretic activity was evaluated by
modified method described by Adams et al
17. Rats were fasted overnight with free access to water before
the experiments. 20% aqueous suspension of Brewer’s yeast (10ml/kg) was
injected subcutaneously in to the animal’s dorsum region to induce
Pyrexia. 17h after the injection, the rectal temperature of each rat was
measured using a Digital Telethermometer. Only rats
that showed an increase in temperature of at least 0.70C
were divided into four groups
of 3 animals each. Group I served as control and received Normal saline
(1ml/kg, p.o). Group II, treated with Aspirin
(300mg/kg, p.o), Group III and IV received MEBM 200 and400
mg/kg, p.o respectively. The
temperature was measured at 1, 2, 3, 5, and 6h after treatment 18,19.
Statistical
Analysis
The statistical significance was determined by using
one way analysis of variance (ANOVA) followed by Dunnet’s
multiple comparison tests. P<0.05 was considered statistically significant.
RESULTS AND DISCUSSION:
The present study revealed that the stem
bark of B.monosperma lam exhibits the Hepatoprotective
and Antipyretic property. The presences of flavonoids,
glycoside, sterols, fixed oils, tannins, were detected on preliminary
phytochemical screening of MEBM. In this study Carbon tetrachloride induced hepato-toxicity model was used to evaluate the hepatoprotective activity. It has been assured that carbon
tetrachloride is the best characterized system of xenobiotic-induced
hepatotoxicity and is frequently used as a model to
study hepatoprotective activity of the drugs 11,12.
CCl4 is metabolised by CYP450 system in
the endoplasmic reticulum to produce trichloromethyl
free radical (CCl3). Trichloromethyl free radical combined with cellular lipids
and proteins in the presence of oxygen which forms trichloromethylperoxyl
radical, which may attack lipids on the membrane of endoplasmic reticulum
faster than trichloromethyl free radical. Thus, trichloromethylperoxyl free radical leads to elicit lipid
per oxidation. The destruction of Ca2+ homeostasis, finally results
in cell death 12,20. This damage to the structural integrity of the
liver is observed from elevated serum levels of hepatospecific
enzymes SGOT, SGPT, ALP and total Bilirubin, and also
decreasing the total protein level. Hepatoprotective
activity of any drug is the ability of its constituents to inhibit the
aromatize activity of CYP450 thereby favouring liver
regeneration 21. The results of CCl4 induced hepato-toxicity were shown in table 1. In control group,
the significant acute hepato cellular damage, and biliary obstruction was indicated by the elevated serum
levels of SGPT, SGOT, ALP, TB and decreased levels of TP. The MEBM (200mg/kg
and 400mg/kg, p.o) significantly (P<0.05)
decreased the elevated serum levels of SGPT, SGOT, ALP, TB and significantly
increased TP levels. These biochemical parameters are comparable with the
standard Liv-52 hepatoprotective drug. Therefore the
MEBM restored the altered levels of enzymes significantly in Carbon tetrachloride induced hepatotoxicity. The results of most several clinical
investigations showed the efficacy and safety of flavonoids
in the hepato- biliary
dysfunction8. Fever may be due to
infection or one of the sequels of tissue damage, inflammation, graft rejection
or other disease states.
