International Research Journal of Plant Science (ISSN: 2141-5447) Vol. 1(3) pp. 062-068, September, 2010
Available online http://www.interesjournals.org/IRJPS
Copyright © 2010 International Research Journals
Full Length Research Paper
Hepatoprotective and antioxidant effects of caesalpinia
bonducella on carbon tetrachloride-induced liver injury
in rats
R. Sambath Kumar1*, K. Asok Kumar1, N. Venkateswara Murthy2.
1 Department of Pharmaceutics and Pharmacology, Faculty of Pharmacy, Seventh of April University, Al-Zawia, Libya.
2. Natural Products Research Laboratory, J. K. K. Nataraja College of Pharmacy, Komarapalayam, 638 183, Namakkal,
Tamilnadu, India.
Accepted 20 September, 2010
The present study was carried out to evaluate the hepatoprotective and antioxidant effect of the
methanol extract of Caesalpinia bonducella (MECB) (Family: Caesalpiniaceae) in Wistar albino rats.
The different groups of animals were administered with carbon tetrachloride (CCl4) (30 % CCl4, 1 ml/kg
b. wt. in liquid paraffin 3 doses (i.p.) at 72 h interval). The MECA at the doses of 50, 100 and 200 mg/kg
and silymarin 25 mg/kg were administered to the CCl4 treated rats. The effect of MECB and silymarin
on serum glutamyl pyruvate transaminase (SGPT), Serum glutamyl oxalacetic acid transaminase
(SGOT) Serum alkaline phosphatase (SALP), bilirubin, uric acid and total protein were measured in the
CCl4 induced hepatotoxicity in rats. Further, the effects of the extract on lipid peroxidation (LPO),
enzymatic antioxidant (superoxide dismutase (SOD) and catalase (CAT)), and non enzymatic
antioxidant (glutathione (GSH), vitamin C and vitamin E) were estimated. The MECB and silymarin
produced significant (p < 0.05) hepatoprotective effect by decreasing the activity of serum enzymes,
bilirubin, uric acid, and lipid peroxidation and significantly (p < 0.05) increased the levels of SOD, CAT,
GSH, vitamin C, vitamin E and protein in a dose dependent manner. From these results, it was
suggested that MECB possess potent hepatoprotective and antioxidant properties.
Key words: Caesalpinia bonducella, hepatoprotective effects, antioxidants, carbon tetrachloride.
INTRODUCTION
Many hepatotoxicants including carbon tetrachloride,
nitrosamines, and polycyclic aromatic hydrocarbons
required metabolic activation, especially by liver
*Corresponding author Email:
Tele: 00218 - 237632147
sambathju2002@yahoo.co.in;
Abbreviations
MECB: methanol extract of Caesalpinia bonducella
LPO: lipid peroxidation
NBT : Nitroblue tetrazolium chloride
DTNB: 5, 5’-dithio bis-2-nitrobenzoic acid
CDNB : 1-Chloro-2,4-dinitrobenzene
TCA : Trichloroacetic acid
cytochrome P450 (P450) enzymes to form reactive, toxic
metabolites that in turn produce liver injury in
experimental animals and humans (Guengerich et al.,
1991) Carbon tetrachloride, a well-known model
compound for the production of chemical hepatic injury,
requires biotransformation by hepatic microsomal P450
to
produce
hepatotoxic
metabolites
namely
trichloromethyl free radicals (CCl.3 and /or CCl3OO-)
(Recknagel et al., 1989). Trichloromethyl free radicals
can react with sulfhydryl groups, such as glutathione
(GSH) and protein thiols, and the covalent binding of
trichloromethyl free radicals to cell proteins is considered
the initial step in a chain of events that eventually lead to
membrane lipid peroxidation and finally to cell necrosis
(Recknagel et al., 1989).
Recently there has been an upsurge of interest in the
Kumar et al. 063
therapeutic potential of medicinal plants as antioxidants
in reducing free radical-induced tissue injury (Pourmorad
et al., 2006). The major constituents of biological
membranes are lipids and proteins. Reactive oxygen
species can easily initiate damage of the cell membrane
constituent that is, phospholipids, and lipoproteins by
propagating a reaction cycle (Raja et al., 2006; Prakash
et al., 2009). It has been mentioned by many authors that
antioxidant activity of plants is due to their phenolic
compounds (Wang, 2003; Wu et al., 2004).
Herbal medicines derived from plant extracts are being
increasingly utilized to treat a wide variety of clinical
diseases, with relatively little knowledge regarding their
modes of action. Caesalpinia bonducella (L.) Flem.,
Fever nut; Bunduc nut, (Family: Caesalpiniaceae)
commonly known as Nata Karanja (Hindi), a prickly shrub
found throughout the hotter parts of India, Myanmar and
Sri Lanka. The chemical constituents of the plant include
flavonoids, triterpenes (Lai et al., 1977; Purushothaman
1982). The leaves of this plant are traditionally used for
the treatment of liver disorders, inflammation and tumors
(Kirtiker and Basu, 1975). It has also been reported to
possess multiple therapeutic properties like antipyretic,
antidiuretic, anthelmintic antibacterial, anticonvulsant,
anti-anaphylactic
and
antidiarrheal,
antiviral,
antiasthmatic, antiamebic and anti-estrogenic. Currently,
the hepatoprotective and antioxidant role of Caesalpinia
bonducella on paracetamol-induced liver damage in rats
(Gupta et al., 2003a) anti-inflammatory, analgesic and
antipyretic activity (Gupta et al., 2003b). Antitumor activity
and antioxidant status of Caesalpinia bonducella against
Ehrlich ascites carcinoma in Swiss albino mice (Gupta et
al., 2004a). Screening of antioxidant and antimicrobial
activities of Caesalpinia bonducella leaves (Gupta et al.,
2004b) were carried out in our laboratory.
Continuing our research the present study was
undertaken to evaluate the protective effects of MECB on
carbon tetrachloride-induced hepatotoxicity. The effect of
MECB on CCl4-induced acute liver injury was also
compared to the effect of silymarin.
Extraction
The dried powder material of the leaf (500 g) was defatted with
petroleum ether (60-800) in a soxhlet apparatus. The defatted
powder material thus obtained was further extracted with methanol
for 72 hours in the soxhlet. The solvent was removed by distillation
under suction and the resulting semi-solid mass was vacuum dried
using rotary flash evaporator to yield (8.78%) a solid residue
(methanol extract). Phytochemical screening of the extract revealed
the presence of alkaloids, saponins, flavonoids, triterpenoids,
tannins, and steroids.
Drugs and chemicals
Silymarin was purchased from Micro labs Hosur Tamilnadu India, 1Chloro-2,4-dinitrobenzene [CDNB], Bovine serum albumin (Sigma
chemical St. Louis, MO, USA), Thiobarbituric acid, Nitroblue
tetrazolium chloride (NBT) (Loba Chemie, Bombay India), 5,5'-dithio
bis-2-nitrobenzoic acid (DTNB), Carbon tetrachloride, (SICCO
research laboratory, Bombay). The solvent and / or reagent
obtained were used as received.
Experimental animals
Studies were carried out using male Wistar albino rats weighing
150–180 g were used. They were obtained from the animal house,
J.K.K.Nataraja College of Pharmacy Tamilnadu, India. The animals
were grouped and housed in polyacrylic cages (38 x 23 x 10 cm)
with not more than six animals per cage and maintained under
standard laboratory conditions (temperature 25 + 2o C) with dark
and light cycle (14/10 h). They were allowed free access to
standard dry pellet diet and water ad libitum. The mice were
acclimatized to laboratory condition for 10 days before
commencement of experiment. All procedures described were
reviewed and approved by the institutional animal ethical
committee.
Toxicity study
For toxicity studies groups of 10 mice were administered (p.o.) with
test compounds in the range of doses 100-1600 mg/kg and the
mortality rates were observed after 72 hours. The LD50 was
determined using the graphical methods of Litchfield and Wilcoxon
(1949).
Experimental studies
MATERIALS AND METHODS
Plant material
The plant grows in all textures of mildly acidic to alkaline soil.
Annual rainfall in the areas where C. bonducella grows in Puerto
Rico ranges from 750 mm to 1800 mm. In India the plants grow
from sea level to 850 m in elevation. The plant Caesalpinia
bonducella was collected from Kolli Hills of Tamilnadu, India. The
plant material was taxonomically identified by the Botanical Survey
of India, Kolkata, India. A Voucher specimen (No.GMS-2) has been
preserved in our laboratory. The leaves were dried under shade
and then powdered with a mechanical grinder and stored in an
airtight container.
Healthy male albino rats were divided into 6 groups each containing
6 animals. Group 1 Normal (Liquid paraffin 1ml/kg body weight,
p.o.) Group 2 (Control) received 30% CCl4 in liquid paraffin (1 ml/kg
body weight, i.p.). Group 3, 4 and 5 received MECB 50, 100 and
200 mg/kg p.o. respectively and Group 6 received standard drug
Silymarin (25 mg/kg p.o) once in a day and CCl4 as mentioned
above. Treatment duration was 10 days and the dose of CCl4 was
administered after every 72-h (Manoj and Aqueed, 2000). Animals
were sacrificed 24 h after the last injection. Blood was collected,
allowed to clot and serum separated. The liver was dissected out
and used for biochemical studies.
Biochemical studies
The blood samples were allowed to clot for 45 min at room
temperature. Serum was separated by centrifugation at 2500 rpm at
064 Int. Res. J. Plant Sci.
30°C for 15 min and utilized for the estimation of various
biochemical parameters like SGPT, SGOT (Bergmeyer et al.,
1978), SALP (King, 1965), serum bilirubin by the method of Malloy
and Evelyn (1937), and protein content was measured by the
method of Lowry et al. (1951). The plasma uric acid was estimated
by the method Caraway (1963).
After collection of blood samples the rats were sacrificed and
their livers excised, rinsed in ice cold normal saline followed by 0.15
M Tris-HCl (pH 7.4) blotted dry and weighed. A 10 % w/v of
homogenate was prepared in 0.15 M Tris-HCl buffer and processed
for the estimation of lipid peroxidation by the method of Ohkawa et
al. (1979). A part of homogenate after precipitating proteins with
Trichloroacetic acid (TCA) was used for estimation of glutathione by
the method of Ellman (1959). Vitamin E by Quaife and Dju, (1948),
with slight modification by Baker and Frank (1951) and Vitamin C by
Omaye et al., (1979) were also estimated. The rest of the
homogenate was centrifuged at 15000 rpm for 15 min at 4° C. The
supernatant thus obtained was used for the estimation of SOD by
the method of Kakkar et al. (1984) and CAT activities were
measured by the method of Aebi (1974).
Statistical Analysis
The experimental results were expressed as mean ± S.E.M. Data
were assessed by using statistical package for social science
(SPSS) version 10.0 using ANOVA followed by Dunnett’s test. P
value of < 0.05 was considered as statistically significant.
RESULTS
Acute toxicity
The methanol extract of leaves of Caesalpinia bonducella
was found to be non-toxic up to doses of 1.6 g/kg and did
not cause any death of the animals tested.
Effect of MECB on serum enzymes, bilirubin, uric
acid and protein
Alteration in the activities of serum enzymes (SGPT,
SGOT and SALP), bilirubin, uric acid and total protein
content in the serum of CCl4 induced liver damage in rats
as evidence from Table1. The level of serum marker
enzymes SGPT, SGOT, SALP, bilirubin and uric acid
were found to be significantly increased and protein
content significantly decreased in CCl4-induced liver
damage rats when compared with the normal group (p <
0.01). Where as treatment with MECB at the dose of 50,
100 and 200 mg/kg showed decreased the activity of
serum transaminase (SGPT, SGOT), ALP, uric acid,
bilirubin and increased the protein content in CCl4induced liver damage in rats compared to that of control
groups (p < 0.05). Silymarin (25 mg/kg) also significantly
decreased the levels of serum enzymes, bilirubin uric
acid and increased the protein content in CCl4 treated
groups as compared with the respective control group.
Effect of MECB on in vivo lipid peroxidation
The localization of radical formation resulting in lipid
peroxidation, measured as MDA (Malondialdehyde) in rat
liver homogenate, is shown in Table 2. MDA content in
the liver homogenate was increased in CCl4 control group
compared to normal group (p < 0.001). MDA level of
MECB 50, 100 and 200 mg/kg group, were inhibited by
49.84, 67.76 and 93.75 % compared to CCl4 control (p <
0.05). At the same time, the effect of silymarin 25 mg/kg
on MDA levels in CCl4 was inhibited by 98.75 %
respectively.
Effect of MECB on SOD and CAT activity in liver
tissues
The effect of MECB on SOD and CAT activities in liver is
shown in Table 2. SOD activity in CCl4 control group was
examined to be lower than in normal group (p < 0.001).
SOD activities in MECB 50, 100 and 200 mg/kg groups
were observed to be higher than in CCl4 control group (p
< 0.05). SOD activities of MECB 50, 100 and 200 mg/kg
were improved by 16.75, 31.80 and 85.12 % respectively.
Silymarin 25 mg/kg also restored the SOD activity in CCl4
treated groups. CAT activity of CCl4 control group was
measured to be strikingly lower than in normal group (p <
0.001). Liver CAT activities in MECB 50, 100 and 200
mg/kg groups were increased by 15.24, 44.35 and 81.34
% respectively when compared with control group (p <
0.05). MECB and silymarin completely restored the
enzyme activity to the normal level at the respective
doses of 200 mg/kg and 25 mg/kg.
Effect of MECB on glutathione, vitamin C and vitamin
E levels in liver tissues
The effect of MECB on glutathione content, vitamin C and
vitamin E levels in the liver is shown in Table 2. GSH
level in normal group was measured to be higher than in
CCl4 control group. GSH level of MECB 50, 100 and 200
mg/kg groups were increased by 12.97, 50.41, and
85.98% respectively as compared to CCl4 control group
(p < 0.05). Silymarin almost completely restored the
glutathione level in CCl4 treated groups to the normal
level. The levels of Vitamin C in the liver of CCl4 control
group decreased in comparison with the normal group (p
< 0.001). After administration of MECB at the dose of 50,
100 and 200 mg/kg., b.w. increased the levels of vitamin
C by 15.78, 47.36 and 77.89% respectively, as compared
to that of the CCl4 control group (p < 0.05). The vitamin E
level in CCl4 control group decreased in comparison with
the normal group (p < 0.001). Treatment with MECB at
the dose of 50, 100 and 200 mg/kg., b.w increased
vitamin E levels by 09.40, 55.85 and 80.85% respectively
when compared to that of the CCl4 control (p < 0.05).
Kumar et al. 065
Table 1. Effect of methanol extract of Caesalpinia bonducella leaves (MECB) on serum enzymes (GPT, GOT and ALP), bilirubin,
protein and uric acid in CCl4 induced hepatic damage in rats
Parameters
Normal (liquid
paraffin 1ml/kg,
b.wt)
Control
(CCl4 1ml/
kg, b.wt)
Silymarin
(25 mg/kg)
+ CCl4
MECB
(50 mg/kg)
+ CCl4
MECB
(100 mg/kg)
+ CCl4
MECB
(200mg/kg)
CCl4
SGOT (U/l)
61.42 ± 3.21
170.21 ± 8.92*
SGPT (U/l)
54.15 ± 2.21
137.51± 7.12*
SALP (U/l)
65.42 ± 3.31
110.55± 5.52*
Bilirubin (mg/dl)
0.98 ± 0.04
2.44 ± 0.15*
Protein (mg/dl)
7.05 ± 0.51
5.25 ± 0.32*
Uric acid (mg/dl)
2.81 ± 0.12
1.36 ± 0.05*
64.32 ± 3.72**
(97.33)
56.41 ± 2.90**
(81.10)
68.94 ± 3.94
(92.28)
1.04 ± 0.04**
(95.89)
6.91± 0.32**
(92.22)
2.76 ± 0.13
(80.00)
157.51± 8.53**
(11.67)
116.15 ± 6.41
(21.36)
98.31± 5.71**
(26.57)
2.23 ± 0.12**
(14.38)
5.93 ± 0.22
(37.77)
2.32 ± 0.11**
(66.20)
102.52 ± 5.81**
(62.22)
83.53 ± 4.42**
(53.98)
84.75± 4.90
(55.44)
1.15 ± 0.05**
(88.37)
6.35± 0.32**
(61.11)
2.48 ± 0.15**
(77.24)
66.41± 3.24**
(95.41)
60.54 ± 2.12**
(71.10)
70.24 ± 3.84**
(89.39)
1.02 ± 0.04**
(97.26)
6.86 ± 0.31**
(89.44)
2.71 ± 0.16**
( 93.10)
+
The data in the parenthesis indicate percent protection in individual biochemical parameters from their elevated values
caused by the CCl4. The % of protection is calculated as 100 X (values of CCl4 control -values of sample) / (values of
CCl4 control - values of vehicle control)
Values are mean ± S.E.M. number of rats=6.
Control group compared with normal group * p < 0.001
Experimental groups compared with CCl4 control group * * p < 0.05
DISCUSSION
It is generally accepted that the hepatotoxicity of
CCl 4 depends on the cleavage of the carbon-chlorine
bond to generate tricloromethyl free radical (.CCl 3 ); this
free radical reacts rapidly with oxygen to form a
trichloromethyl peroxy radical (.CCl 3 O2 ). This metabolite
may attack membrane polyunsaturated fatty acids and
causes lipid peroxidation which plays a main role in the
induction of liver injury (Agarwal et al., 2006) and further
causes impairment of membrane function. CCl 4 -induced
hepatic injuries are commonly used as models for the
screening of hepatoprotective plant extract and the extent
of hepatic damage is assessed by the level of released
cytosolic transaminases including SGPT and SGOT in
circulation (Starzynska et al., 2008) When administrated
methanol extract exhibited protection against CCl 4 induced liver injuries as manifested by the reduction of
toxin-mediated rise in serum enzymes in rats. Our results
using the model of CCl4-induced hepatotoxicity in the rats
demonstrated that MECB at the different doses caused
significant inhibition of SGPT and SGOT levels. Serum
ALP and bilirubin levels on other hand are related to the
function of hepatic cell. Increase in serum level of ALP is
due to increased synthesis, in presence of increasing
biliary pressure (Moss and Butterworth, 1974). Our
results using the model of CCl4-induced hepatotoxicity in
rats demonstrated that MECB at different doses caused
significant inhibition of SALP and bilirubin levels. Effective
control of bilirubin level and alkaline phosphates activity
points towards an early improvement in the secretory
mechanism of the hepatic cell.
Uric acid, the metabolic end product of purine
metabolism, has proven to be a selective antioxidant,
capable especially of reacting with free radicals and
hypochlorous acid (Hasugawa and Kuroda, 1989). The
reduced level of uric acid in hepatotoxicity conditions may
be due to the increased utilization of uric acid against
increased production of the free radicals, which is a
characteristic feature of cancer condition. The reversal of
altered uric acid level to near normal in MECB treated
rats could be due to strong antioxidant property of MECB,
which contributes to its antioxidant potency.
Liver cell injury induced by CCl4 involves initially the
metabolism of CCl4 to trichloromethyl free radical by the
mixed-function oxidase system of the endoplasmic
reticulam. It is postulated that secondary mechanisms
link CCl4 metabolism to the widespread disturbances in
hepatocyte function. These secondary mechanisms could
involve the generation of toxic products arising directly
from CCl4 metabolism or from peroxidative degeneration
of membrane lipids (Halliwell et al., 1992). In our study,
elevations in the levels of end products of lipid
peroxidation in liver of rat treated with CCl4 were
observed. The increase in MDA level in liver suggests
enhanced lipid peroxidation leading to tissue damage and
066 Int. Res. J. Plant Sci.
Table 2. Effect of the methanol extract of Caesalpinia bonducella leaves (MECB) on lipid peroxidation (LPO) antioxidant enzymes
(SOD and CAT) and non enzymatic antioxidant (GSH, vitamin E and vitamin C) in the liver of CCl4 intoxicated rats.
Parameters
Normal (liquid
paraffin
1ml/kg, b.wt)
Control
1ml/
kg, b.wt)
Lipid peroxidation
(n
mole
of
MDA/mg
protein)
Glutathione
content
(µg/mg protein)
0.92 ± 0.05
Vitamin E (mg/g/wet
tissue)
Vitamin C (mg/g/wet
tissue)
Superoxide dismutase
(U/mg protein)
Catalase
(U/mg protein)
(CCl4
Silymarin
(25 mg/kg)
+ CCl4
MECB
(50 mg/kg)
+ CCl4
MECB
(100 mg/kg)
+ CCl4
MECB
(200mg/kg) + CCl4
7.45 ± 0.31*
1.13 ± 0.05
(98.75)
4.26 ± 0.21**
(49.84)
3.12 ± 0.12 (67.76)
1.45 ± 0.06**(93.75)
5.45 ± 0.29
0.67 ± 0.32*
5.20 ± 0.24
(94.76)
1.29 ± 0.06**
(12.97)
3.08 ± 0.19**(50.41)
4.78 ± 0.25** (85.98)
4.12 ± 0.21
2.24 ± 0.14*
3.76 ± 0.15** (80.85)
0.61± 0.02*
1.06 ± 0.06**(47.36)
1.35 ± 0.07** ( 77.89)
93.36 ± 5.35
52.23 ± 2.26*
65.31 ± 3.29 (31.80)
87.24 ± 4.32 (85.12)
354.61±
15.07
267.92 ±
12.07*
2.41 ± 0.10**
(09.40)
0.76 ± 0.02**
(15.78)
59.12 ± 2.23**
(16.75)
281.14 ± 15.03**
( 15.24)
3.29 ± 0.11**(55.85)
1.56 ± 0.03
3.98 ± 0.15
(92.55)
1.43 ± 0.04
(86.36)
90.56 ± 4.43
(93.19)
353.63 ± 15.25
(98.86)
306.37 ± 13.09**
(44.35)
338.41 ± 16.05**(81.31)
The data in the parenthesis indicate percent protection in individual biochemical parameters from their elevated values
caused by the CCl4. The % of protection is calculated as 100 X (values of CCl4 control -values of sample) / (values of
CCl4 control - values of vehicle control)
Values are mean ± S.E.M. number of rats=6. Control group compared with normal group * P < 0.001
Experimental groups compared with CCl4 control group * * P < 0.05
failure of antioxidant defense mechanisms to prevent
formation of excessive free radicals. Treatment with
MECB significantly reversed these changes. Hence it
may be possible that the mechanism of hepatoprotection
of MECB is due to its antioxidant effect.
The role of free radical reactions in disease pathology
is well established, suggesting that these reactions are
necessary for normal metabolism but can be detrimental
to health; the antioxidants protected against free radicals
induced oxidative damage by antioxidant enzymes such
as superoxide dismutase and catalase or antioxidant
compounds (Kakoti et al., 2007).
Biological systems
protect themselves against the damaging effects of
activated species by several means. These include free
radical scavengers and chain reaction terminators;
enzymes such as SOD, CAT and GPx system. (Kurata et
al., 1993). The SOD dismutates superoxide radicals O2into H2O2 plus O2, thus participating, with other
antioxidant enzymes, in the enzymatic defense against
oxygen toxicity. In this study, SOD plays an important
role in the elimination of ROS derived from the
peroxidative process of xenobiotics in liver tissues. The
observed increase of SOD activity suggests that the
MECB have an efficient protective mechanism in
response to ROS. And also, these findings indicate that
MECB may be associated with decreased oxidative
stress and free radical-mediated tissue damage.
CAT is a key component of the antioxidant defense
system. Inhibition of these protective mechanisms results
in enhanced sensitivity to free radical-induced cellular
damage. Excessive production of free radicals may result
in alterations in the biological activity of cellular
macromolecules. Therefore the reduction in the activity of
these enzymes may result in a number of deleterious
effects due to the accumulation of superoxide radicals
and hydrogen peroxide. Administration of MECA
increases the activities of catalase in CCl4 induced liver
damage rats to prevent the accumulation of excessive
free radicals and protects the liver from CCl4 intoxication.
GSH is a naturally occuring substance that is abundant
in many living creatures. It is widely known that a
deficiency of GSH within living organisms can lead to
tissue disorder and injury. For example, liver injury
included by consuming alcohol or by taking drugs like
acetaminophen, lung injury by smoking and muscle injury
by intense physical activity (Leeuwenburgh et al., 1995),
all are known to be correlated with low tissue levels of
GSH. From this point of view, exogeneous MECB
supplementation might provide a mean of recover
reduced GSH levels and to prevent tissue disorders and
injuries. The present study, we have demonstrated the
effectiveness of MECB by using CCl4 induced rats, which
are known models for both hepatic GSH depletion and
injury.
The availability of vitamin C is a determined factor in
controlling and potentiating many aspects of host
resistance to cancer. The decreased level of vitamin C
was found CCl4 control animals. Vitamin C can protect
cell membranes and lipoprotein particles from oxidative
damage by regenerating the antioxidant form of vitamin E
Kumar et al. 067
(Das, 1994). Thus, vitamin C and E act synergistically in
scavenging a wide variety of ROS. The recoupment of
vitamin C to near normal level in drug treated rats was
found to be due to the potent antioxidant activity of bark
extract which may induce the regeneration of ascorbic
acid. Vitamin E is the most significant antioxidant of its
kind in animal cells and it can protect against chemical
carcinogenesis and tumor growth (Buettner, 1993).
Significantly decreased vitamin E levels in CCl4 control
animals might be due to the excessive utilization of this
antioxidant for quenching enormous free radicals
produced in these conditions. The increased level of
vitamin E in drug treated rats reveals the antioxidative
nature of the MECB. The extract may scavenge the free
radicals and thus maintains the normal level of vitamin E.
It has been reported that Caesalpinia. bonducella
contain flavonoid and triterpenoid (Lai et al., 1977;
Purushothaman 1982). A number of scientific reports
indicated certain flavonoids, triterpenoids and steroids
have protective effect on liver due to its antioxidant
properties (DeFeudis, 2003; Takeoka, and Dao, 2003).
Presence of those compounds in MECB may be
responsible for the protective effect on CCl4 induced liver
damage in rats.
In conclusion, the results of this study demonstrate that
MECB has a potent hepatoprotective action upon carbon
tetrachloride-induced hepatic damage in rats. Our results
show that the hepatoprotective and antioxidant effects of
MECB may be due to its antioxidant and free radical
scavenging properties. Further, investigation is underway
to determine the exact phytoconstituents that is
responsible for its hepatoprotective and antioxidant
activity.
ACKNOWLEDGEMENT
One of the author Ramanathan Sambath Kumar, grateful
to AICTE, New Delhi, India, for providing financial support
to this work. The authors gratefully acknowledge our
beloved Secretary & correspondent N. Sendamaraai
J.K.K. Nataraja Educational Institution Komarapalayam,
Tamilnadu, India, for provided us the abound facilities.
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