GARCINIA CAMBOGIA AND ETHANOL INDUCED PEROXIDATION

Indian Journal of Pharmacology 2001; 33: 87-91 SHORT COMMUNICATION

Correspondence: C.S. Shyamala Devi
THE MODULATING EFFECT OF GARCINIA CAMBOGIA EXTRACT ON ETHANOL
INDUCED PEROXIDATIVE DAMAGE IN RATS
P. MAHENDRAN, C.S. SHYAMALA DEVI
Department of Biochemistry & Molecular Biology, University of Madras, Guindy Campus,
Chennai - 600 025.
Manuscript Received: 25.5.2000 Revised: 14.6.2000 Accepted: 13.7.2000
Objective: To determine the modulating effect of Garcinia cambogia fruit extract on ethanol induced
peroxidative damage in rats.
Method: Male albino rats weighing 125 to 150g were administered ethanol (7.11g per kg body weight /
day) for 45 days. Ethanol administered rats were treated concomitantly with Garcinia cambogia fruit
extract (1g/kg body weight / day) orally for 45 days. After the experimental period the antioxidant enzymes,
LPO, conjugated diene in the liver tissue, serum AST, ALT and alkaline phosphatase and lipid levels in
both serum and liver tissue were estimated.
Results: Co-treatment of the rats with Garcinia cambogia significantly inhibited the rise in lipid levels
and also the peroxidative damage caused by ethanol, which is evident from the improved antioxidant
status. The levels of serum AST, ALT and alkaline phosphatase were maintained at near normalcy in
Garcinia cambogia treated rats.
Conclusion: The imbalance in lipid metabolism could be the reason for increase in lipid peroxidation. In
our present study the treatment with Garcinia cambogia fruit extract resulted in reduction of both serum
and liver lipid to near normalcy. This hypolipidemic property of Garcinia cambogia in turn reduces the
peroxidative damage, enhanced by ethanol.
Ethanol Garcinia cambogia lipid peroxidation hyperlipidemia antioxidant enzymes
SUMMARY
KEY WORDS
INTRODUCTION
Liver being the major site for detoxification is the pri-
mary target for environmental or occupational toxic
exposure
1
. The alcoholic liver injury appears to be
generated by the effects of ethanol metabolism and
the toxic effects of acetaldehyde which may be me-
diated by acetaldehyde altered proteins
2
. Chronic
alcohol intake is known to produce hypercholestero-
lemia, hyperlipidemia, hypertriglyceridemia
3,4
. In
chronic lipid accumulation the liver cells become fi-
brotic and leads to impaired liver function. Enhanced
lipid peroxidation has been reported in hyper-
lipidemia
5
, which is also induced by ethanol
3,4
. Etha-
nol increases triglycerides and cholesterol levels thus
inducing imbalance in lipid metabolism in liver, heart,
kidney and other organs and this could explain the
reason for the increase in lipid peroxidation in these
organs. Recently free radical induced lipid peroxida-
tion has gained much importance because of its in-
volvement in several pathologies
6,7
. Protection of cell
membrane from lipid peroxidation becomes a neces-
sity to prevent, cure or delay the aforesaid patho-logies.
Rind of the fruits of Garcinia cambogia (Gaertn.)
Desr. (Clusiaceae) is an astringent and is useful in
the treatment of ulcers, haemorrhoids, diarrhoea and
dysentry
8,9
. Garcinia cambogia fruit extract, contain-
i ng the pri nci pl e organi c aci d, (-)-erythro-L
s
-
hydroxycitric acid is a powerful anti-lipogenic agent
10
.
The present study is an attempt to elucidate the
antiperoxidative property of Garcinia cambogia on
ethanol induced peroxidative damage and its efficacy
to inhibit lipid peroxidation.
MATERIALS AND METHODS
Animals: Male albino rats weighing 125 to 150 g
were purchased from FIPPAT, Padappai, Chennai.
The animals were housed in plastic bottom cages
P. MAHENDRAN AND C.S. SHYAMALA DEVI
Table 1. Serum and hepatic levels of total cholesterol, triglycerides, free fatty acids, phospholipids, alkaline phosphatase, aspartate
amino transferase (AST), alanine amino transferase (ALT) in experimental groups
a
.
I II III IV ANOVA
Parameters
b
Control Group 18% Ethanol treated Garcinia cambogia Garcinia cambogia F value*
(n = 6) group (n = 6) treated group + 18% ethanol treated
(n = 6) group (n = 6)
Total cholesterol A 70.4 1.18 113.2 2.19*** 64.3 1.26*** 72.2 1.81*** 4.24
B 4.36 0.37 6.82 0. 51*** 3.72 0.51* 4.76 0.40*** 2.88
Triglycerides A 107.8 2.08 156.0 2.14*** 98.5 2.09*** 112.9 2.11*** 3.97
B 4.23 0.41 5.87 0.50*** 3.73 0.46* 4.54 0.40*** 2.94
FFA A 64.1 1.96 115.4 2.28*** 56.6 1.64*** 70.8 2.07*** 3.68
B 11.5 0.93 21.6 1.67*** 9.9 0.81** 13.2 1.24*** 3.24
Phospholipids A 115.6 2.06 142.4 2.17*** 110.3 1.98** 126.7 2.23*** 3.07
B 23.1 2.15 38.7 1.83*** 20.6 1.91* 30.4 1.47*** 2.90
Alkaline Phosphatase A 0.76 0.05 1.57 0.04*** 0.69 0.04
NS
0.86 0.03*** 2.78
B 1.41 0.04 0.88 0.03*** 1.34 0.05
NS
1.19 0.04*** 2.91
AST A 0.48 0.04 0.83 0.03*** 0.41 0.04
NS
0.52 0.04*** 2.83
B 0.72 0.06 0.46 0.04*** 0.78 0.05
NS
0.69 0.05*** 2.79
ALT A 0.63 0.04 0.92 0.04*** 0.61 0.04
NS
0.58 0.03*** 2.96
B 1.04 0.03 0.68 0.02*** 1.07 0.03
NS
0.93 0.03*** 2.81
A : Serum B : Liver.
a
Values are expressed as mean SD for 6 animals in each group.
Students t test: Group II vs Group I, Group III vs Group I, Group IV vs Group II. *p < 0.05; **p < 0.01; ***p < 0.001
b
The values of lipids in serum are expressed as mg/dl. The values of AST, ALT and ALP in serum are expressed as kat/litre.
The levels of AST and ALT in liver are expressed as moles of pyruvate liberated/sec/g protein. The level of ALP in liver is expressed
as moles of phenol liberated/sec/g protein.
F - test, * Significant at level of 5% (2.77).
and allowed free access to standard laboratory chow
(Hindustan Lever Foods, Bangalore, India) and water.
Drug: Garcinia cambogia fruit extract was obtained
from Siris Herbex, Vijayawada, India. Absolute alco-
hol (99%) was purchased from Anilax Chemicals,
USA. All other chemicals used for the experiment
were of analytical grade.
Grouping
Group I - Normal Control (n = 6).
Group II - Rats given 18% ethanol (7.11g/kg body
weight day) 5 ml/100 g body weight for
45 days (n = 6).
Group III - Normal rats given Garcinia cambogia
1 g/kg body weight/day orally for 45 days
(n = 6).
Group IV - Rats given 18% ethanol (7.11g/kg/body
weight/day) + Garcinia cambogia, 1 g/
kg body weight/day for 45 days (n = 6).
After the experimental period the overnight fasted
rats were sacrificed by cervical dislocation. Blood and
tissues were collected in ice-cold containers for vari-
ous estimations.
AST, ALT
11
and alkaline phosphatase
11
were esti-
mated in the serum. SOD
12
, CAT
13
, GSH
14
, GSH-Px
15
,
GST
16
, LPO
17
and conjugated dienes
18
were esti-
mated i n the l i ver ti ssue. Total chol esterol
19
phospholipids
20,21
, triglycerides
22
FFA
23
and protein
24
were estimated both in serum and tissue.
88
GARCINIA CAMBOGIA AND ETHANOL INDUCED PEROXIDATION
Table 2. The level of lipid peroxide, conjugated diene and activities of antioxidant enzymes in the liver of experimental groups
a
.
I II III IV ANOVA
Parameters
b
Conrol group 18% Ethanol Garcinia cambogia Garcinia cambogia + F value*
(n = 6) treated group treated group 18% ethanol treated
(n = 6) (n = 6) group (n = 6)
SOD 59.8 2.56 26.4 0.98*** 62.2 2.34
NS
54.4 1.76*** 3.68
CAT 72.6 3.14 44.5 3.52*** 70.8 2.26
NS
66.7 2.82*** 3.20
LPO 3.87 0.18 7.21 0.10*** 3.70 0.12
NS
4.06 0.09*** 2.79
Conjugated diene 74.8 0.69 98.3 1.14*** 73.6 0.75
NS
81.2 0.97*** 3.54
GST 152.6 3.46 81.8 2.85*** 155.4 3.71
NS
151.3 2.97*** 4.12
GSH-px 175.2 6.82 102.7 5.84*** 182.3 7.06
NS
171.8 5.44*** 3.96
GSH 5.11 0.05 2.16 0.08*** 4.89 0.09
NS
4.32 0.04*** 2.84
a
Values are expressed as mean SD for 6 animals in each group
Students t test: Group II vs Group I, Group III vs Group I, Group IV vs Group II. ***p<0.001
b
Lipid peroxide (LPO) = moles of TBA reactants/g protein. Conjugated diene = mM/100g wet tissue; Superoxide dismutase (SOD) =
kat/g protein. Catalase (CAT) = nmoles of H
2
O
2
decomposed/sec/g protein. Glutathione peroxidase (GSH - Px) = mmoles of GSH
utilised/sec/g protein, Glutathione-S-transferase (GST) = moles of CDNB conjugated/sec/g protein. Glutathione (GSH) = nmoles of
GSH/g tissue.
F - test, * Significant at level of 5% (2.77).
RESULTS
The results are summarised in Table 1 and 2. The diet
intake of rats in different groups were almost similar.
After 45 days, the weight gain of ethanol fed rats was
less than that of normal control rats. The activity of
AST, ALT, and alkaline phosphatase increased signifi-
cantly (p<0.001) in the serum after 45 days in ethanol
treated rats (Table 1). The activity of superoxide
dismutase catalase, GSH, GST, GSH-Px in the liver
decreased significantly (p<0.001) in alcoholic rats
(Table 2). The alcohol treated rats caused a signifi-
cant increase (p<0.001) in the LPO content and con-
jugated diene in the liver when compared with that of
normal rats. Concentration of cholesterol, triglyceride,
FFA and phospholipids showed significant (P<0.001)
increase in serum and liver tissue in alcohol treated
rats. Normal rats treated with Garcinia cambogia
(Group III) showed a decrease in lipid levels alone
both in serum (p<0.001) and liver tissue (p<0.05).
DISCUSSION
Lipid peroxidation is a complex and natural deleteri-
ous process. The effect of free radicals on human
beings have recently been considered as their close
relation to toxicity and diseases
25
. Chronic adminis-
tration of ethanol was found to produce an accumu-
lation
4
of cholesterol and triglycerides in the liver as
well as in the blood. This imbalance in lipid metabo-
lism could play a role in aggravating the lipid peroxi-
dation
5
. Varga et al, have proven that hyperlipidemia
and elevated lipid peroxidation are inter-related
26
. The
possible explanation could be the hyperlipidemic con-
dition would stimulate the catabolic pathway via
oxidative breakdown
5
.
HMG CoA reductase catalyses the rate limiting step
in cholesterol biosynthesis and its activity directly
correlates with the extent of tissue cholesterol syn-
thesis which is significantly activated during ethanol
ingestion
27
. Plasma LCAT is the enzyme involved in
89
P. MAHENDRAN AND C.S. SHYAMALA DEVI
the esterification of cholesterol in the plasma. In-
crease in serum cholesterol, in the present study in
alcohol treated rats may possibly be the result of
decreased activity of this enzyme
27
.
Increase in serum triglycerides in alcohol treated rats
may be due to decreased activity of lipoprotein li-
pase which is involved in the uptake of triglyceride
rich lipoprotein by extra hepatic tissue. Increased
synthesis or decreased lipid deposition or both re-
sulted in simultaneous accumulation of lipids in the
blood and in the liver
27
.
Ethanol induces hyperlipidemia
3,4
and hyperlipidemia
enhances lipid peroxidation
5
causing hepatotoxicity by
increasing the free radical formation which in turn in-
creases the level of lipid peroxides in hepatic tissue.
Garcinia cambogia containing the principal organic
acid (-)-erythro-L
s
-hydroxycitric acid is an effective
inhibitor of ATP-citrate lyase which cleaves citrate to
produce acetyl CoA.
10
(-)-hydroxycitrate suppresses
biosynthesis of both fatty acids and cholesterol in rat
hepatocytes
28,29
and in rat liver,
30
hence cholesterol
and triglyceride levels are drastically reduced in
Garcinia cambogia treated experimental rats (Group
IV). The reduction in lipids indirectly lowers the level
of peroxides due to hyperlipidemia.
The two antiperoxidative enzymes namely SOD and
CAT decreased significantly in the hepatic tissues of
alcohol administered rats suggesting the increased
damage to this tissue as a result of uncontrolled gen-
eration of partially reduced oxygen species. The lev-
els of AST, ALT and alkaline phosphatase in the se-
rum have been elevated in ethanol treated rats sug-
gesting hepatotoxicity as a result of high ethanol in-
take
31
.
Garcinia cambogia, an effective antilipogenic agent
32
,
prevents the hepatic cells to become fibrotic and the
cellular damage due to hyperlipidemia. This is evi-
dent from the near normal activities of transferases
and ALP in the liver of Garcinia cambogia treated
group (Table 1).
Glutathione protects the hepatocytes by combining
with the reactive metabolites and thereby preventing
their covalent binding to liver protein
31
. Liver glutath-
ione after alcohol administration was found to de-
crease due to increased utilization by the hepato-
cytes because GSH seems to act as scavengers for
toxic chemical agents. The non availability of glutath-
ione decreases the activity of glutathione peroxidase
and glutathione transferase. Glutathione acts as the
substrate for both GSH-Px and GST. Depletion of
glutathione will render the enzymes (GSH-Px & GST)
inactive and/or less active.
It was established that the content of primary
(acylhydroperoxide) and secondary (intermolecular
seams in aminophospholipids) products of lipid
peroxide oxidation in blood of patients with the back-
ground of hyperlipidemia and hypercholesterolemia
increased intensively
33
. Also in our present study the
levels of LPO and conjugated dienes increased sig-
nificantly in the ethanol administered rats suggest-
ing its pathogenic role. Treatment of rats with Garcinia
cambogia inhibited the deleterious process of lipid
peroxidation and maintained the levels of glutathione
to near normalcy.
The observed abnormalities in the liver and serum
may be in part due to hyperlipidemia leading to
changes in the activity of antiperoxidative enzymes,
glutathione and increased lipid peroxidation. Garcinia
cambogia modulates its antiperoxidative role by in-
hi bi ti ng hyperl i pi demi a and peroxi dati on of
biomembranes which is sufficient to cause cell death
when uncontrolled.
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