Life Sciences 76 (2004) 585 – 597

www.elsevier.com/locate/lifescie

Protective effect of Centella asiatica on antioxidant tissue defense

system against adriamycin induced cardiomyopathy in rats
A. Gnanapragasama, K. Kumar Ebenezara, V. Sathisha, P. Govindarajub, T. Devakia,*
a
Department of Biochemistry, University of Madras, Guindy Campus, Chennai- 600 025, Tamilnadu, India
b
Department of Soil Chemistry, Agriculture Research Institute, Madurai, Tamilnadu, India
Received 24 May 2004; accepted 4 August 2004

Abstract

Increased oxidative stress and antioxidant deficit have been suggested to play a major role in adriamycin
induced cardiomyopathy and congestive heart failure due to multiple treatments with adriamycin. In this study the
cardio protective effect of Centella asiatica on myocardial marker enzymes and antioxidant enzymes in
adriamycin induced cardiomyopathy was investigated in rats. The rats administered with adriamycin (2.5 mg/kg
body wt, i.p) caused myocardial damage that was manifested by the elevation of serum marker (LDH, CPK, GOT
and GPT) enzymes and showed significant changes in the antioxidant enzymes (SOD, CAT, GPx, GST). Pre-co-
treatment with Centella asiatica(200 mg/kg of body wt/oral) extract significantly prevented these alterations and
restored the enzyme activities to near normal levels. These findings demonstrate the cardio protective effect of
Centella asiatica on antioxidant tissue defense system during adriamycin induced cardiac damage in rats.
D 2004 Elsevier Inc. All rights reserved.

Keywords: Centella asiatica; Adriamycin; Cardiomyopathy; Antioxidant enzymes; Lipid peroxidation

Introduction

Adriamycin, also known as doxorubicin, is a potent antitumor antibiotic used for the treatment of a
variety of soft and solid human malignancies. However, treatment may be complicated by its acute and
chronic side effects. One major chronic side effect is the development of cardiomyopathy and ultimately

* Corresponding author. Tel.: +91 44 22351269; fax: +91 44 22352494.

E-mail address: devakit@yahoo.co.uk (T. Devaki).

0024-3205/$ - see front matter D 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.lfs.2004.09.009
586 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597

congestive heart failure (Buja et al., 1973; Lefrak et al., 1973; Singal et al., 1995). Adriamycin was
originally isolated from a mutant Streptomyces peucetius obtained from the daunorubicin producing
organism, S. Peucetius.
Several different mechanisms that have been suggested for adriamycin induced cardiomyopathy are:
inhibition of nucleic acid and protein synthesis (Buja et al., 1973); release of vaso active amines (Bristow
et al., 1980); changes in adrenergic function (Tong et al., 1991); lysosomal alterations (Singal et al.,
1985); free radical formation and oxidative stress (Bristow et al., 1980; Doroshow, 1983; Kalyanaraman
et al., 1980; Singal et al., 1987); reduction in myocardial antioxidants (Siveski-Iliskovic et al., 1994;
Revis and Marusic, 1978) and lipid peroxidation (Singal et al., 1987; Myers et al., 1977). It appears that
oxygen radical induced injury is involved in most of the above proposed mechanisms.
Medicinal plants play a key role in human health care now-a-days. While in acute conditions such as
myocardial necrosis the modern medicine is the only answer, it is increasingly being understood that the
traditional Indian medicine does have a remedy for these acute disorders.
Centella asiatica (L.) urban (umbelliferae) is commonly used in the ayurvedic system of medicine to
treat various diseases. The plant is claimed to posses anti-inflammatory (Suguna et al., 1996), memory
improvement (Veerendrakumar and Gupta, 2002) and anticancer activity (Babu et al., 1995). Centella
asiatica is also useful in vascular diseases such as venous hypertension and atherosclerosis (De Sanctis
et al., 2001; Incandela et al., 2001). Based on the hypothesis of free radical mediated toxicity, Centella
asiatica has been recently reported to have anti-lipid peroxidative (Katare and Ganachari, 2001) and free
radical scavenging activities (Jayashree et al., 2003).
The active constituents present in the centella asiatica extract are triterpenes namely asiatic acid,
asiaticoside (Inamdar et al., 1996). Also, different parts of Centella asiatica were found to contain high
phenolic contents (3.23–11.7 g/100 g dry sample), which exhibit strong association with its antioxidative
activities (Zainol et al., 2003).
Hence, the present investigation was undertaken to study the activities of markers of cardiac functions
and antioxidant enzymes during adriamycin induced cardiomyopathy and the effect of aqueous extract of
Centella asiatica pre-co-treatment in reducing the extent of damage in the myocardium.

Materials and methods

Chemicals

Adriamycin was procured from Dabur Pharmaceuticals (Doxorubicin Hydrocholoride-Adrim), New

Delhi, India. All other chemicals used were of analytical grade.

Plant materials and preparation of extract

Plants were collected in and around the campus of University of Madras, Chennai, Tamil Nadu,India.
It was authenticated at the herbarium of Botany, Center for Advanced Studies in Botany, University of
Madras. The whole plant was shade dried and coarsely ground with grinder. The coarse powder of plant
was extracted with 8 parts of distilled water under boiling for 5 hrs and was filtered through a 400-mesh
cloth to collect the extract. The extract was concentrated and freeze dried to get a powder of greenish
brown in colour (Gupta et al., 2003).
 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597 587

Animals

Adult male albino rats of Wistar strain weighing about 140–160 g were obtained from the Tamilnadu
Veterinary and Animal Science University, Chennai, India. They were acclimatized to animal house
conditions, fed commercial pelleted rat chow (Hindustan Lever Ltd., Bangalore, India) and had free
access to water. This study was conducted according to the ethical norms approved by Ministry of Social
Justices and Empowerment, Government of India and by Animal Ethics Committee Guidelines of our
institution.

Experimental design

The rats were divided into four groups of six animals in each group as follows: Group 1, Control;
Group 2, Adriamycin administered; Group 3, Centella asiatica extract administered; Group 4, Centella
asiatica treated plus adriamycin administered. Drug administration was as follows: Centella asiatica
extract was given orally (200 mg/kg body wt) through an intragastric feeding tube over a period of three
weeks, one week prior to the adriamycin administration and two weeks along with adriamycin
administration. This particular dosage was fixed after trying out different doses for different days in the
same set of rats. The dosage having optimum protective effect was chosen for the study. Adriamycin was
given intraperitoneally (2.5 mg/kg body weight) in six equal injections over a period of two weeks for a
cumulative dose of 15 mg/kg body weight (Siveski-Iliskovic et al., 1994, 1995).
At the end of the experimental period, the body weight was determined and rats were sacrificed by
cervical decapitation. Blood was collected and the separated serum was used for further estimation. The
heart was excised immediately, rinsed in ice-cold saline, dried, weighed and homogenized in Tris-HCl
buffer of pH 7.4 (0.1 M) using a Teflon homogenizer. The tissue homogenate was then centrifuged in a
cooling centrifuge at 500 g to remove the debris and the supernatant was used for the analysis of
biochemical parameters. The tissue homogenate was placed at –20 8C until further use. All the
experiments were carried out in a dark cold room at 4 8C.

Biochemical parameters

Estimation of Protein

Protein was estimated by the method of Lowry et al. (1951). To 0.1 ml of homogenate or serum, 0.9
ml of water and 4.5 ml of alkaline copper reagent were added and kept at room temperature for 10 min.
Then 0.5 ml of Folin’s reagent was added and the colour developed was read at 640 nm after 20 min. The
level of protein was expressed as mg/g of tissue or mg/dl of serum.

Lactate dehydrogenase (LDH)

Lactate dehydrogenase was assayed according to the method of King (1965). To 1.0 ml of the
buffered substrate, 0.1 ml of enzyme preparation was added and the tubes were incubated at 37 8 C for
15 min. After adding 0.2 ml of NAD+ solution, the incubation was continued for another 15 min. The
reaction was arrested by adding 0.1 ml of DNPH (2,4-dinitrophenyl hydrazine), and the tubes were
588 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597

incubated for a further period of 15 min at 37 8C after which 7.0 ml of 0.4 NaOH (sodium hydroxide)
solution was added and the colour developed was measured at 420 nm in a Shimadzu UV
spectrophotometer. Suitable aliquots of the standards were also analyzed by the same procedure. The
activity of the enzyme was expressed as Amoles of pyruvate liberated/mg protein/hour.

Creatine Phosphokinase (CPK)

Serum creatine phosphokinase activity was determined by the method of Okinaka et al. (1961). The
reaction mixture comprised of 0.05 ml of serum, 0.1 ml of substrate, 0.1 ml of ATP solution and 0.1 ml
of cysteine-hydrochloride solution. The final volume was made up to 2.0 ml with distilled water and
incubated at 37 8C for 30 min. The reaction was arrested by the addition of 1.0 ml of 10% TCA
(Trichloroacetic acid) and the contents were subjected to centrifugation. To 0.1 ml of the supernatant, 4.3
ml distilled water and 1.0 ml ammonium molybdate were added and incubated at room temperature for
10 min. 0.4 ml of ANSA was added and the colour developed was read at 640 nm after 20 min. The
activity of the enzyme was expressed as Amoles of phosphorus liberated/mg protein/hour.

Glutamate oxaloacetate transaminase (GOT)

The activity of glutamate oxaloacetate transaminase was assayed by the method of Bergmeyer and
Bernt (1974). To 1.0 ml of substrate, 0.2 ml of serum was added and incubated for 1 hr at 37 8C. Then
1.0 ml of 0.02% DNPH was added and kept at room temperature for 20 min. To the control tube, sample
was added after arresting the reaction with DNPH. Then 5 ml of 0.4 N NaOH was added and the colour
developed was read at 540 nm. The activity was expressed as Amoles of pyruvate liberated/mg protein/
hour.

Glutamate pyruvate transaminase (GPT)

Glutamate pyruvate transaminase was assayed by the method of Bergmeyer and Bernt (1974). The
assay mixture containing 1.0 ml of substrate and 0.2 ml of serum was incubated for 1 hr at 37 8C. To the
control tubes, sample was added after the reaction was arrested by the addition of 1.0 ml DNPH. The
tubes were kept at room temperature for 30 min. Then 5.0 ml of 0.4 N NaOH was added and the colour
developed was read at 540 nm. The activity was expressed as Amoles of pyruvate liberated/mg protein/
hour.

Total Reduced Glutathione (GSH)

Reduced glutathione was estimated by the method of Ellman (1959). 0.1 ml of tissue homogenate
was precipitated with 5% TCA. The contents were mixed well for complete precipitation of proteins
and centrifuged. To 0.1 ml of supernatant, 2.0 ml of 0.6 mM DTNB [5,5 dithiobis(2-nitrobenzoic
acid)] reagent and 0.2 M phosphate buffer (pH 8.0) were added to make up to a final volume of 4.0
ml. The absorbance was read at 412 nm against a blank containing TCA instead of sample. A series of
standards treated in a similar way also run to determine the glutathione content. The amount of
glutathione is expressed as nmoles/g heart tissue. 5-sulphosalicylic acid was used to prevent the
oxidation of glutathione.
 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597 589

Glutathione peroxidase (GPx)

GPx was assayed by the method of Rotruck et al. (1973). The reaction mixture consisted of 0.2 ml of
0.8 mM EDTA, 0.1 ml of 10 mM sodium azide, 0.1 ml of 2.5 mM H2O2, 0.2 ml of reduced glutathione,
0.4 ml of 0.4 M phosphate buffer pH 7.0, and 0.2 ml homogenate and was incubated at 37 8C for 10 min.
The reaction was arrested by the addition of 0.5 ml of 10% TCA and the tubes were centrifuged at 2000
rpm. To the supernatant 3.0 ml of 0.3 mM disodium hydrogen phosphate and 1.0 ml of 0.04 % DTNB
were added and the colour developed was read at 420 nm immediately. The activity of GPx was
expressed as Amoles of glutathione oxidized/min/mg protein.

Glutathione S-transferase (GST)

GST was assayed by the method of Habig et al. (1974). To 0.1 ml of homogenate, 1.0 ml of 0.3 M
phosphate buffer pH 6.5, 1.7 ml of water and 0.1 ml of 30 mM CDNB (1-chloro-2, 4-dinitrobenzene)
were added. After incubation at 37 8C for 15 min, 0.1 ml of GSH was added and change in OD was read
at 340 nm for 3 min at an interval of 30 sec. Reaction mixture without the enzyme was used as blank.
The glutathione-S-transferase activity was expressed as units/min/mg protein.

Superoxide dismutase (SOD)

SOD was assayed by the method of Misra and Fridovich (1972). 0.1 ml of tissue homogenate
was added to the tubes containing 0.75 ml ethanol and 0.15 ml chloroform (chilled in ice) and
centrifuged. To 0.5 ml of supernatant, added 0.5 ml of 0.6 mM EDTA solution and 1 ml of 0.1 M
carbonate-bicarbonate (pH 10.2) buffer. The reaction was initiated by the addition of 0.5 ml of 1.8
mM epinephrine (freshly prepared) and the increase in absorbance at 480 nm was measured in a
Shimadzu UV spectrophotometer. The enzyme activity is expressed as 50% inhibition of epinephrine
autooxidation.

Catalase (CAT)

CAT was assayed by the method of Takahara et al. (1960). To 1.2 ml of 50 mM phosphate buffer pH
7.0, 0.2 ml of the tissue homogenate was added and reaction was started by the addition of 1.0 ml of 30
mM H2O2 solution. The decrease in absorbance was measured at 240 nm at 30 sec intervals for 3 min.
The enzyme blank was run simultaneously with 1.0 ml of distilled water instead of hydrogen peroxide.
The enzyme activity was expressed as Amoles of H2O2 decomposed/ min/mg protein.

Myocardial lipid peroxides (LPO)

Lipid peroxidation (LPO) was assayed by the method of Ohkawa et al. (1979) in which the
malondialdehyde (MDA) released served as the index of LPO. 1,1,3,3-Tetra ethoxypropane
malondialdehyde bis (diethyl acetal), was used as standard. To 0.2 ml of homogenate, 0.2 ml of 8.1%
SDS, 1.5 ml of 20% acetic acid (pH 3.5) and 1.5 ml of 0.8% TBA were added. The mixture was made up
to 4 ml with water and then heated in a water bath at 95 8C for 60 min using glass ball as a condenser.
After cooling, 1 ml of water and 5 ml of n-butanol/pyridine mixture were added and shaken vigorously.
590 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597

After centrifugation at 4000 rpm for 10 min, the organic layer was taken and its absorbance was
measured at 532 nm. The level of lipid peroxides was expressed as nmoles of MDA formed / mg of
protein.

Statistical analysis

All the grouped data were statistically evaluated with SPSS/10 software. Hypothesis testing methods
included one way analysis of variance (ANOVA) followed by least significant difference (LSD) test. P
values of less than 0.05 were considered to indicate statistical significance. All these results were
expressed as Mean F S.D for six animals in each group.

Results

General observations

Figs. 1 and 2 shows the body and heart weight of the control and experimental groups of rats
respectively. Body and heart weight were significantly (p b 0.05) lowered in group 2, adriamycin
intoxicated rats. Pre-co treatment with Centella asiatica extract (group 4) resulted in a significant
increase in the body and heart weight gain when compared to adriamycin intoxicated rats. This marked
gain in the body and heart weight of Centella asiatica treated rats was nearer to that of the control rats
(Group 1).

Cardiac marker enzymes

Table 1 shows the activities of marker enzymes such as LDH, CPK, GOT and GPT in the serum of
control and experimental groups of rats. Marked elevation (p b 0.05) in the activities of these enzymes
were observed in group 2, adriamycin intoxicated rats when compared with group 1, control rats.
Activities of these enzymes in serum were maintained at near normal (p b 0.05) levels in the group 4

Fig. 1. Effect of Centella asiatica on the body weight of control and experimental groups of rats.
 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597 591

Fig. 2. Effect of Centella asiatica on the heart weight of control and experimental groups of rats. Results are expressed as
mean F SD (n= 6). *p b 0.05. Comparisons are made between group 1 (Control) with group 2 (Adriamycin induced) and
group 2 with group 4 (Centella asiatica + adriamycin).

rats, pre-co-treated with Centella asiatica extract. Group 3, rats administered with Centella asiatica
extract alone did not show any changes when compared to group 1, control rats.

Myocardial antioxidants

Table 2 presents the level of GSH and the activities of GPx, GST, SOD and CAT in the heart of
control and experimental groups of rats. A significant (p b 0.05) decrease was observed in GSH content
and in the activities of antioxidant enzymes GPx, GST, SOD and CAT (p b 0.05) in the group 2,
adriamycin intoxicated rats. Pre-co-treatment with Centella asiaticaextract (group 4) significantly
prevented (p b 0.05) these alterations when compared to group 2, adriamycin intoxicated rats.

Myocardial lipid peroxides

Fig. 3 indicates the level of lipid peroxides (LPO) in the heart of control and experimental groups of
rats. Maximum induction of LPO was noticed in group 2, adriamycin intoxicated rats when compared to

Table 1
Effect of Centella asiatica on the activities of marker enzymes in serum of control and experimental groups of rats
Groups (Treatment) LDH CPK SGOT SGPT
1. Control 1089.91 F 94.30 83.05 F 8.56 79.05 F 8.96 23.25 F 1.64
2. Adriamycin induced 1653.71 F 140.24* 189.68 F 18.84* 154.39 F 15.39* 42.94 F 2.86*
3. Centella asiatica alone 1090.68 F 92.02 83.13 F 7.15 79.46 F 8.07 24.43 F 1.39
4. Centella asiatica + adriamycin 1160.85 F 99.60* 92.05 F 8.38* 86.83 F 8.60* 29.90 F 2.12*
Results are expressed as mean F S.D (n= 6).
Comparisons are made between group 1 (Control) with group 2 (Adriamycin induced) and group 2 with group 4 (Centella
asiatica + adriamycin).
Activity is expressed as: Amole of pyruvate liberated/mg of protein/hr for LDH, GOT and GPT; Amol of phosphorus liberated/
mg protein/hr for CPK.
* p b 0.05.
592 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597

Table 2
Effect of Centella asiatica on antioxidant enzymes in the heart of control and experimental groups of rats
Group (Treatment) GSH GPx GST SOD CAT
1. Control 2.75 F 0.21 1.77 F 0.21 0.32 F 0.03 8.95 F 0.85 86.80 F 10.38
2. Adriamycin induced 1.29 F 0.13* 1.20 F 0.11* 0.19 F 0.02* 4.70 F 0.36* 61.42 F 5.52*
3. Centella asiatica alone 2.66 F 0.21 1.59 F 0.20 0.30 F 0.02 9.58 F 0.97 89.40 F 8.49
4. Centella asiatica + Adriamycin 2.69 F 0.22* 1.74 F 0.14* 0.34 F 0.02* 8.19 F 0.85* 85.55 F 10.33*
Results are expressed as mean F SD (n = 6).
Comparisons are made between group 1 (Control) with group 2 (Adriamycin induced) and group 2 with group 4 (Centella
asiatica + adriamycin).
Activity is expressed as: Unit/min/mg protein for GST; Amole of GSH oxidized /min/mg protein for GPx; nmole/g heart tissue
for GSH; 50% inhibition of epinephrine autoxidation for SOD; Amole of H2O2 decomposed/min/mg protein for CAT.
* p b 0.05.

group 1, control rats. The altered metabolic changes were significantly (p b 0.05) restored to near normal
levels in the (group 4) rats treated with Centella asiatica extract.

Discussion

Adriamycin induced cardiomyopathy is a very well documented phenomenon in humans (Lefrak et

al., 1973; Singal and Iliskovic, 1998) as well as in different animal models (Siveski-Iliskovic et al., 1994;
Myers et al., 1977; Jones et al., 1990; Chalcroft et al., 1973). Studies have shown that the adriamycin
toxicity is proceeding via the production of free radicals (Doroshow, 1983; Bristow et al., 1980) and are
characterized by hypotension, tachycardia and various arrhythmias. The reduction in body weight was
attributed to reduced food intake and inhibition of protein synthesis due to adriamycin toxicity (Tong et
al., 1991). Treatment with Centella asiatica extract prevents the changes in body weight as well as
development of congestive heart failure.

Fig. 3. Level of lipid peroxides in heart of control and experiemental groups of rats. Results are expressed as mean F SD (n= 6).
*p b 0.05. Comparisons are made between group 1 (Control) with group 2 (Adriamycin induced) and group 2 with group 4
(Centella asiatica + adriamycin).
 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597 593

Increased activity of serum LDH, CPK, GOT and GPT is a well-known diagnostic marker of
myocardial function. It has been reported that these enzymes are released from the heart into blood
stream, thus increasing their concentration in serum (Wexler and Kittinger, 1963; Sathish et al., 2002;
Deepa and Varalakshmi, 2003). In the present study, marked elevation in the activities of these
enzymes in the serum of adriamycin intoxicated rats were observed. Pre-co-treatment with aqueous
extract of Centella asiatica resulted in significant reduction in the levels of these enzymes towards
near normal as compared with cardiomyopathy-induced rats. This establishes that the active ingredient
triterpenes (asiatic acid, asiaticoside) may be responsible for the cardio protective effect of centella
asiatica extract. It has been reported that another structurally related triterpene (arjunolic acid) possess
the cardio protective effect by inhibition of enzymatic activity against isoproterenol induced
myocardial damage (Sumitra et al., 2001). Antioxidants constitute the foremost defense system that
limit the toxicity associated with free radicals. It has been demonstrated that free radicals are produced
by adriamycin (Singal et al., 1987; Iliskovic et al., 1999; Doroshow, 1983; Kalyanaraman et al., 1980)
and that antioxidant enzymes play a critical role in detoxifying free radicals (Kaul et al., 1993).
Several studies have been reported that antioxidant enzyme activities are reduced by multiple
adriamycin treatments in different animal species (Revis and Marusic, 1978; Doroshow et al., 1980;
Iliskovic et al., 1999).
Free radical scavenging enzymes such as catalase, superoxide dismutase, glutathione peroxidase and
glutathione-S-transferease are the first line of cellular defense against oxidative injury. The equilibrium
between these enzymes is an important process for the effective removal of oxygen stress in intracellular
organelles (Andrew and Mathew, 1989).
GSH functions as a free radical scavenger in the repair of radical induced cellular damage. Low level
of GSH was observed during increase in oxidative stress caused by adriamycin administration (Iliskovic
et al., 1998). This observation supports our findings where we have observed a decline in GSH level in
adriamycin intoxicated rats. Oral administration of aqueous extract of Centella asiatica along with
adriamycin intoxicated rats maintained the concentration of GSH at near control levels.
It has been suggested that GPx plays an important role in protecting the heart from peroxidative
attack (Doroshow et al., 1980). GPx activity is significantly depressed by the repeated treatment with
adriamycin in a sub-chronic study (Siveski-Iliskovic et al., 1994, 1995). The decreased activity of GPx
and GST were observed in adriamycin intoxicated rats, which may be due to the reduced availability
of GSH. The increased level of glutathione in turn enhances the activities of GPx and GST and thus
helps in scavenging the free radicals. Increase in the activities of GPx and GST upon oral
administration of aqueous extract of Centella asiatica in adriamycin intoxicated (group 4) rats shows
the antioxidant potential of the plant extract against injury caused by free radical species (Jayashree et
al., 2003).
The cytoprotective action of Centella asiatica, which brings about mitigation of peroxide production,
paves the way for the reversal of the reduced antioxidant potential of the cells. This was in accordance
with the observation that triterpenes exhibit antiperoxidation activity against oxalate induced toxicity
(Vidhya et al., 2000; Sasa et al., 1994).
SOD and CAT are important antioxidant enzymes in mitigating free radical induced cell injury. A
decrease in the activity of superoxide dismutase and catalase could result in the decreased removal of
superoxide ion and hydrogen peroxide radicals which brings about a number of reactions which are
harmful to the myocardium (Sumitra et al., 2001). Decrease in the activities of these enzymes could be due
to increased reactive oxygen free radicals, which can themselves reduce the activity of these enzymes
594 A. Gnanapragasam et al. / Life Sciences 76 (2004) 585–597

(Hodgson and Fridovich, 1975; Searle and Willson, 1980). This is in agreement with our findings where
adriamycin intoxicated rats showed decreased activities of SOD and CAT. The rats pre-co-treated with
aqueous extract of Centella asiatica and adriamycin showed increased activities of these enzymes which
suggest that the extract may have ability to prevent the deleterious effects induced by free radicals. It has
been reported that Centella asiatica is a free radical scavenger and its supplementation increases the
activities of SOD and CAT (Jayashree et al., 2003). This forms the basis for the cardio protective effect of
Centella asiatica.
Adriamycin can undergo a two-electron reduction to form alkylating quinone and methides
(Kalyanaraman et al., 1980). As quinones, they are also able to undergo redox cycling to generate
oxygen radicals, which can in turn lead to the induction of lipid peroxidation. Increased levels of oxygen
species due to adriamycin have been detected by an increase in tissue malondialdehyde formation, which
is a breakdown product of lipid peroxidation (Myers et al., 1977; Singal et al., 1987). Significant
elevation in the level of LPO after adriamycin administration was observed in this study. The rats
administered aqueous extract of Centella asiatica orally plus adriamycin showed a significant decrease
lipid peroxidation status when compared with adriamycin intoxicated rats. This might be due to the anti-
lipidperoxidative property of Centella asiatica(Katare and Ganachari, 2001) that protects the
myocardium from lipid peroxidation. The previous study also revealed that the Centella asiatica have
suppressive effect on myloperoxidase activity, which in turn may decrease the production of oxygen
species and reduce concomitant tissue damage (Cheng et al., 2004).
The cardio protective effect of Centella asiatica may be due to reduction of hydroperoxides,
inactivation of free radicals, chelation of metals or combinations thereof (Zainol et al., 2003). Previous
studies suggested that chelating the metals has met with some success in reducing adriamycin induced
cardiomyopathy (Singal et al., 2000). In further, it may suggest that most of these biological effect of
Centella asiatica may be related to presence of active ingredients such as triterpenes and phenolic
compounds. The cardio protective and antioxidant activity of these compounds are well documented
(Sumitra et al., 2001; Somova et al., 2003; Shukla et al., 1999; Cook and Samman, 1996) against free
radical induced cell damage.

Conclusion

In conclusion, the present study implies that Centella asiatica may be a particularly useful agent as it
could enhance myocardial antioxidants and significantly prevent the heart from adriamycin induced
oxidative stress. Therefore, it could offer a useful support to the adriamycin therapy by acting as a cardio
protective agent and thus prevents the extent of cardiac damage. Further studies are in progress to
elucidate the exact mechanisms of action of the various constituents in Centella asiatica and their
potential in the treatment and/or prevention of cardio toxicity.

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