evaluation of hepatoprotective activity of bauhinia purpurea linn.pdf

Vol. 2 (3) Jul – Sep 2011 www.ijrpbsonline.com 1389
International Journal of Research in Pharmaceutical and Biomedical Sciences ISSN: 2229-3701
_________________________________________Research Paper
Evaluation of Hepatoprotective Activity of Bauhinia purpurea Linn
Veena Rani I1*, Veena G1, Bhagavan Raju M2, Tejeswini G1, Uday Bhasker G1 and
Sowmya P1
1Department of Pharmacology, CM College of Pharmacy, Maisammaguda, Dhulapally,
Hyderabad, Andhra Pradesh, India.
2Department of Pharmaceutical Chemistry, CM College of Pharmacy, Maisammaguda,
Dhulapally, Hyderabad, Andhra Pradesh, India.
__________________________________________________________________________________
ABSTRACT
Present study was carried out to investigate different extracts of Bauhinia purpurea (B.P) for its
hepatoprotective activity against CCl4 induced hepatotoxicity. Mature leaves of Bauhinia purpurea were
collected, authenticated and was subjected to extraction using different solvents like chloroform, alcohol and
water. Healthy wistar albino rats (150-200g) of male sex were used for the in-vivo investigations. Liver damage
was induced by administration of 30% CCl4 suspended in olive oil (1ml/kg body weight). Activities of liver
marker enzymes, glutamate oxaloacetate transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT),
acid phosphatase (AP), alkaline phosphatase (ALP),total albumin(TA), total bilirubin(TB), Total protein(TP),
direct bilirubin (DB) at a dose of aqueous extract of leaves (100 mg/kg) chloroform extract of leaf of B.P
(100mg/kg and 150 mg/kg) and ethanol extract of leaf of B.P (100mg/kg and 150 mg/kg) showed a significant
hepatoprotective effect in comparison with the standard (sylimarin). It is also confirmed by liver
histopathology of treated animals. The present study demonstrated the extracts of B.P have hepatoprotective
effect against CCl4 induced hepatotoxicity.
Key Words: Bauhinia purpurea, CCl4 and hepatoprotective.
INTRODUCTION
Abundant natural substances, inorganic chemicals
which are absorbed through intestinal tract, nasal
route or by parenteral route are metabolized by
liver. Cytochrome P450 reductase enzyme system
which is present in the smooth endoplasmic
reticulum of liver facilitates the metabolism of
diverse substances. Over dosage or excess use of
the drugs, chemical agents, microcystins etc
generally referred as hepatotoxins which cause
hepatotoxicity1
. Hepatotoxicity is the main reason
for most of the drugs to be withdrawn from market.
Liver is an adaptable organ of the body that
regulates the physiological environment. Liver
injuries have been a major toxicological problem in
the course of treatment for chronic ailments like
tuberculosis, cancer, hepatitits etc. A publication
report revealed that isolation of flavonids like
flavonolignan named as silymarin from the plant
Silybum marianum showed hepatoprotective
activity. From this discovery many researchers paid
attention world wide on medicinal plants for
________________________________________
*Address for correspondence:
E-mail: vina913@gmail.com
hepatoprotective activity2
.
The toxicity induced by CCl4 may be attributed to
oxidative stress which is indicated by considerable
elevation in the biochemical parameters like
SGOT, SGPT, ALP AP etc. ALP serves as a
specific marker for hepatic damage as it is present
richly in liver cells3
.
SGOT found principally in the cytoplasm of the
liver cells serves as a measure of the integrity of the
hepatocytes, and correlates to the number of
hepatocytes exaggerated. The leakage of the
enzyme into the blood stream indicates the injury to
the hepatocytes4
.SGPT virtually found in almost
every tissue of the body is useful for the production
of antioxidnats in conditions of oxidative stress5
.
HEPATOTOXICITY
Various factors like plant products, minerals, and
also some products of bacterial and fungal
metabolism lead to hepatotoxicity. Some of the
pharmaceutical, chemical products and the waste
materials are hepatotoxins which enter into the
human body6
.
One of the potent hepatotoxic agents is CCl4, when
it is ingested it gets circulated to all organs because

of its high lipid solubility. On ingestion of toxic
doses it causes blockage in the lipoprotein
synthesis and produces fat accumulation in the liver
because these lipoproteins carry the triglycerides
away from this organ. The synthesis of proteins is
reduced and there is a rapid decline in the levels of
cytochrome P450 as well as glucose 6-phosphatase
and the sequestration of Ca2+
ions by Ca2+
ATPase
is reduced by the endoplasmic reticulum, as a result
there is an increased intracellular Ca2+
concentration7
.
Now a days liver disease has become a serious
health hazard. Herbs play an imperative role in the
treatment liver diseases due to lack of consistent
liver shielding drugs in allopathic therapeutic
practices8
.Liver is considered vital organ as it
determines the nutritional plane by its processes.
The action of toxins, pathogens and poisons can be
antagonized by liver tonics and helps the liver cells
to reduce inflammation and stimulates
regeneration capacity of liver9
Bauhinia purpurea L. is a medium sized deciduous
tree which belongs to the family Cesalpiniaceae10
.
Phytochemical study revealed the presence of
glycosides, saponins, flavonoids, phenolic
compounds, triterpenoids, oxepins, fatty acids and
phytosterols11
. B. purpurea was used to cure
various diseases in several ayurvedic traditional
medicine systems. Several activities like
antidiabetic, analgesic, antibacterial, anti-diarrheal,
anti-inflammatory, nephroprotective, wound
healing activity, antioxidant activity, cardiac
activity, antimalarial, antimycobacterial, antifungal,
anticancerous, and thyroid hormone regulating
activity has been reported12-22
.
Therefore the study has been proposed to evaluate
the in-vivo hepatoprotective activity of B.P extracts
based on the fact reported earlier on decreased
invitro lipid peroxidation.
MATERIALS AND METHODS
Drugs & Chemicals
Silymarin is obtained from SIGMA
CHEMICALS, MUMBAI, CCl4 was procured
from S.D FINE Chem. Ltd, MUMBAI.
Suspensions of Bauhinia purpurea extracts were
prepared in 1% tween 80. All chemicals used in the
study were obtained chemically and were of
analytical grade.
Preparation of Plant extract
The aqueous, alcoholic and chloroform extracts of
B.P were prepared by maceration and continuous
soxhlet extraction respectively. The leaves of the
authenticated whole plant (Voucher no. 0215) were
collected from Forest academy, Government of
Andhra Pradesh in the month of December. The
leaves were shade dried at room temperature,
powdered and sieved through no.44 size. The
aqueous extract was obtained by macerating 100gm
of the powder with 500ml of double distilled water
for 24h with intermittent shaking ( yield = 5%w/w).
The alcoholic and chloroform extracts were
obtained by hot soxhlet extraction with 100gm of
powdered drug with 500ml with 3cycles (yield- 2%
w/w).
EXPERIMENTAL ANIMALS
Male Wistar albino rats weighing about (150-200
g) were used. The animals were housed in
polypropylene cages and maintained at 27o
 2o
C at
12 h light dark cycle. They were acclimatized to the
laboratory conditions for 5 days prior to use in
animal house (reference number 1217/A/08
CPCSEA). The animals were fed with standard
laboratory feed and drinking water ad libitum.
Protocol
Eight groups consisting of 6 animals each were
used for the study. Group I served as control which
received 1% tween 80 solution by I.P.route. Group
II were given Slylimarin ( 100mg/kg I.P.), Group
III with CCl4 in olive oil (0.5ml/kg I.P.), Group IV
and V received alcoholic extract (100 and 150
mg/kg I.P. respectively), Group VI and VII were
given chlorofrom extract ( 100 and 150 mg/kg I.P.
respectively). Group VIII was given aqueous
extract (100mg/kg I.P). All the doses were
administered for a period of 10 days with CCl4
from 4th
day to 10th
day of the study. The blood
sample was collected on 11th
day from the
retroorbital plexus with ether anesthesia.
In-vivo hepatoprotective activity
The in-vivo hepatoprotective activity was evaluated
on the basis of biochemical parameters viz., SGOT,
SGPT, serum albumin and total protein levels,
alkaline and acid phosphatase levels, and bilirubin
levels and histopathology of the liver tissues. All
the serum enzymes were analysed using kits
available commercially.
The isolated liver tissues were suspended in 10%
formalin and were sent for evaluation of liver
toxicity by histopathological studies at National
Institute of Nutrition (CSIR), Hyderabad. The
paraffin fixed liver tissue samples were then
subjected to photomicroscopic observations. The
results were presented in figure 1-8
Statistical analysis
All the values are represented as Mean  SEM.
Statistical significance were analyzed using
ANOVA followed by Dunnett’s T test using Based
fitting program (Prism, Graph pad). p values <0.05
were considered significant

RESULTS
Biochemical parameters
The aqueous extract showed a decrease in the
levels of ALP, ASP, TB and DB (p <0.001) and
SGOT (p <0.01), SGPT, TP and AP (p <0.05)
Table no: 1 & 2 when compared to standard
sylmarin. The alcoholic extracts showed a dose
dependent decrease in the enzyme levels of ALP,
ASP, SGOT, SGPT (p <0.001).TP, AP (p <0.01),
DB (p <0.05) and TB (p <0.001). The chloroform
extracts showed a dose dependent decrease in the
enzyme levels of ALP, ASP, SGOT, SGPT (p
<0.001).TP, DB, AP (p <0.01), and TB (p <0.05)
The histopathology of the liver samples revealed a
significant dose dependent protection by the
extracts against CCl4 induced hepatotoxicity. The
fatty degeneration, lymphocytic infiltration,
steatosis and necrosis from ballooning degeneration
to necrosis changes were analysed. The maximum
protection was seen with alcoholic and chloroform
extracts 150 mg/kg I.P. and standard sylmarin
(100mg/kg I.P.).
DISCUSSION
The present study revealed a significant decrease in
the serum enzyme levels which can be attributed to
hepatoprotection. BP extract was found to decrease
the levels of ALP, ACP significantly and there is a
dose dependent decrease in the elevated SGOT and
SGPT levels of the extracts when compared to CCl4
group.
CCl4 treated Liver showed perivenular necrosis,
steatosis with degree of steatosis being variable
from ballooning degeneration to necrosis. Central
lobular vacuoles, frequently dilated and congested
central veins were seen with dilatation of
surrounding sinusoids, which contradicted to the
observations of standard sylmarin, the aqueous,
chloroform and alcoholic extracts showed a clear
portal tract and central vein with normal lobular
architecture and decreased cell degeneration
indicating the hepatoprotective action of extracts of
B.purpurea. The histopathological studies further
confirmed the above results presented in fig 1-8.
Therefore, from the above study the extracts of
Bauhinia purpurea exhibited potent
hepatoprotective activity against CCl4 induced liver
toxicity which can be ascribed to its ability to
decrease the oxidative damage.
Table 1: Biochemical Parameters of extracts of B.Purpurea against CCl4 induced hepatotoxicity
Values are expressed as Mean ± SEM; * p<0.05; ** p< 0.01; *** p<0.001 when compared with standard (Silymarin)
Table 2: Biochemical Parameters of extracts of B.Purpurea against CCl4, induced hepatotoxicity
Values are expressed as Mean ± SEM; * p<0.05; ** p< 0.01; *** p<0.001 when compared with standard (Silymarin)
GROUPS ALP SGOT SGPT ALBUMIN
Control 109±1.167***
29.13±0.28***
35.75±1.153**
2.3±0.186***
Silymarin (100mg/kg) 126±1.202***
43.35±0.33***
44.55±0.6102***
3.383±0.20***
CCl4 (0.5ml/kg) 270±4.551***
251.2±6.43***
279.8±4.430***
2.017±0.1***
Alcoholic extract (100mg)+
CCl4
163.7±2.3***
63.57±0.46***
74.67±1.022***
1.7±0.058***
Alcoholic extract (150mg) +
CCl4
153.2±3.02***
58.53±0.53***
63.32±0.773***
1.48±0.05***
Chloroform Extract (100mg)+
CCl4
179.5±2.49***
61.9±0.368***
76.5±0.7632***
1.67±0.1***
Chloroform Extract (150mg)+
CCl4
163±1.673***
57.45±0.43***
59.29±0.461***
1.48±0.13***
Aqueous extract + CCl4 187±3.022***
55.0±1.317**
52.5±0.7638*
1.78±0.08***
GROUPS TP TB DB AP
Control 4.72±.125*
0.0298±0.008***
0.183±.0033*
14.33±0.760***
Silymarin (100mg/kg) 5.32±0.61***
0.128±.003***
0.151±.00715***
19.83±0.477***
CCl4 (0.5ml/kg) 6.53±0.31***
0.246±0.009***
0.187±0.0067**
24.33±0.4216**
Alcoholic extract (100mg)+
CCl4
4.57±.0843**
0.158±0.002***
0.12±.0.0577*
15.00±1.655**
Alcoholic extract (150mg) +
CCl4
4.42±0.833***
0.153±0.001**
0.117±.00601**
12.00±1.265***
Chloroform Extract
(100mg)+ CCl4
4.53±0.154**
0.18±0.0024***
0.1167±0.0033**
15.67±0.4216**
Chloroform Extract
(150mg)+ CCl4
4.38±0.138***
0.175±0.0013***
.02±.00472***
9.0±0.3651***
Aqueous extract + CCl4 4.63±.0955*
0.173±0.004***
0.06±.07342***
23.17±0.5426*

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