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Abstract. In the present study, the cardioprotective action of tender coconut water (TCW) in
experimental myocardial infarction induced by isoproterenol in rats was studied. The results
indicated that feeding TCW afforded protection against induction of myocardial infarction.
There was decreased concentration of total cholesterol, VLDL + LDL- cholesterol and HDL-
cholesterol in the serum of isoproterenol treated rats fed TCW. The cholesterol levels in
the heart and aorta were also lower in these groups. Triglycerides and phospholipids in the
serum, heart and aorta were lower in isoproterenol treated rats given TCW. Recovery from
myocardial damages was evident from the values of marker enzymes viz creatine phospho-
kinase (CPK), serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate
transaminase (SGPT) and lactate dehydrogenase (LDH). Induction of myocardial infarction in
rats by isoproterenol administration resulted in increased activities of SGOT, SGPT and LDH
in the serum and heart and CPK in the serum. On the other hand, isoproterenol treated rats
fed TCW showed decreased activities of these enzymes. Histopathological studies showed
very little myocardial damage in isoproterenol treated rats fed TCW. The observed beneficial
effects of TCW may be due to several factors viz potassium, calcium, magnesium, L-arginine
that are present in the water.
Introduction
coronary heart disease [5]. In addition, it is a major source of the free amino
acid L-arginine, which is reported to have cardioprotective action [6]. Thus
the present study was done to investigate the effect of TCW in experimentally
induced myocardial infarction in rats.
of lipids and for the assay of enzymes. For histopathological examination, the
heart was removed and fixed in 10% buffered neutral formalin.
Biochemical analysis. Lipids in the serum and tissues were extracted by the
method of Folch et al. [9]. Aliquots of the organic extracts were evaporated
to dryness and used for the estimation of cholesterol [10], triglyceride [11],
phospholipids [12], and serum lipoprotein cholesterol [13].
Activity of creatine phosphokinase (CPK) was measured using a reagent
kit obtained from TECO DIAGNOSTICS, USA. Serum glutamate oxalo-
acetate transaminase (SGOT) and serum glutamate pyruvate transaminase
(SGPT) were measured by the method of Reitman et al. [14]; lactate dehyd-
rogenase (LDH) by the method of King [15]. Protein content was estimated
by the method of Lowry et al. [16] after TCA precipitation.
Statistical analysis. The SPSS statistical program was employed. The res-
ults were evaluated using analysis of variance (ANOVA) utilizing the F test.
The results are presented as the mean value ± SD for the control and ex-
perimental rats. Differences among the means for the groups were assessed
using the Duncans Multiple Range Test to determine which mean values were
significantly different at p < 0.05.
Results
In the present study, the food intake, body weight gain and food efficiency
ratio of control and TCW fed rats did not differ significantly.The food intake
per day of control rats was 12 ± 2 g and TCW fed rats was 12.5 ± 2.1 g, the
food efficiency ratio (diet / weight gain) was 6.30 and 6.26 respectively. The
weight gain of control rats and TCW fed rats were 66.6 ± 4.1 g and 67.0 ±
4.5 g.The survival rate on induction of myocardial infarction by isoproterenol
was significantly higher in rats fed TCW when compared to control rats. The
survival rate was 91% in the TCW fed group and 66% in the control group.
Induction of myocardial infarction in untreated rats by isoproterenol ad-
ministration resulted in increased activities of serum CPK, SGOT, SGPT and
LDH compared to normal rats (Table 1). The activities of SGOT,SGPT and
LDH in the heart were similar to those in the serum (Table 2). On the other
hand, isoproterenol treated rats fed TCW had decreased activities of the en-
zymes in the serum (Table 1). The activities were also decreased in the hearts
of the rats given TCW (Table 2).
Isoproterenol alone treated rats (Group 1 b) had elevated concentrations
of total cholesterol, very low density lipoprotein (VLDL) + low density lipo-
protein (LDL) – cholesterol and high density lipoprotein (HDL) – cholesterol
in the serum compared with the control (Group 1a) (Figure 1). These con-
centrations were significantly lower in isoproterenol treated rats given TCW
(Group 2 b). The cholesterol levels in the heart and aorta were also lower in
this group (Figure 2).
The concentrations of triglycerides and phospholipids in the serum, heart
and aorta were higher in isoproterenol alone treated rats (Group 1 b) when
compared to normal rats (Group 1 a) (Figure 3 and 4). On the other hand iso-
proterenol treated rats fed TCW (Group 2 b) had lower levels of triglycerides
and phospholipids in the serum, heart and aorta (Figure 3 and 4). Rats given
isoproterenol alone (Groups 1 b) had focal areas of degeneration, with loss of
sacroplasm and infiltration with mononuclear cells and necrosis in the heart
while isoproterenol treated rats fed TCW (Group 2 b) had very little evidence
of myocardial necrosis, and the heart was found to be almost normal. Rats
given TCW alone (Group 2 a) showed no perceptible changes in the heart
when compared to control rats (Figure 5).
Discussion
Figure 2. Concentration of cholesterol in heart and aorta of animals given isoproterenol and
TCW (mg/100 gm tissue).
∗ Indicates that the result is significantly different from group 1a.
Indicates that the result is significantly different from group 1b.
Significance was accepted at p < 0.05.
TCW = Tender Coconut Water.
CPK as well as the higher activities of SGPT, SGOT and LDH in the blood
and heart are indicative of myocardial damage. Isoproterenol is known to
cause myocardial infarction and it may increase serum enzymes [19]. In ad-
dition, the concentrations of cholesterol, triglycerides and phospholipids also
increased in the serum, heart and aorta. Higher levels of serum and tissue
lipids have been reported by others [20, 21]. That isoproterenol administered
rats had severe myocardial damage was evident from the histopathological
studies. Feeding TCW afforded protection against induction of myocardial
infarction. Decreased concentration of cholesterol, triglycerides and phos-
pholipids in TCW treated rats indicated the beneficial effects of TCW in
Figure 3. Concentration of triglycerides in seurm, heart and aorta of animals given isoproter-
enol and TCW (mg/100 ml and mg/100 gm tissue).
∗ Indicates that the result is significantly different from group 1a.
Indicates that the result is significantly different from group 1b.
Significance was accepted at p < 0.05.
TCW = Tender Coconut Water.
Figure 4. Concentration of phospholipids in serum, heart and aorta of animals given isoproter-
enol and TCW (mg/100 ml and mg/100 gm tissue).
∗ Indicates that the result is significantly different from group 1a.
Indicates that the result is significantly different from group 1b.
Significance was accepted at p < 0.05.
TCW = Tender Coconut Water.
Figure 5. Cross sections of normal and experimental hearts of rats stained with hematoxylin-eosin × 100. A – Control, B – Control + isoproterenol, C
– TCW, D – Isoproterenol + TCW.
TCW = Tender Coconut Water.
10
References
1. Pradera ES, Fernander E, Calderin O (1942) Coconut water – A clinical and experi-
mental study. Am J Dis Child 64: 977–996.
2. Shaw M, Srivastava BIS (1963) Purine like substances from coconut endosperm and
their effect on senescence in excised cereal leaves. Pl Physiol 38: 528–531.
3. Adams W, Bralt DE (1992) Young coconut water for home rehydration in children with
mild gastroenteritis. Trop Geogr Med 44: 149–153.
4. Macalalag EV, Macalalag AL (1987) Bukolysis: young coconut water renoclysis for
urinary stone dissolution. Int Surg 72: 247.
5. Anonymous (1961) Biochemical composition of coconut water. Curr Sci 30: 363.
6. Cooke IP, Singer AH, Tsao P, Zera P, Roman RA, Billinghan MP (1992) Antiatherogenic
effects of L-arginine in hypercholesterolemic rabbit. J Clin Invest 90: 1168–1171.
7. Guidlines on the care of laboratory animals and their use for scientific purposes (1987)
University Federation For Animal Welfare: Hearts, United Kingdom.
8. Sheela Sasikumar C, Shyamala Devi CS (2000) Effect of Abana an ayurvedic formula-
tion of lipid peroxidation in experimental myocardial infarction in rats. Ind J Exp Bio
38: 827–830.
9. Folch J, Less M, Sloane-Stanley GH (1957) A simple method for the isolation and
purification of total lipids from animal tissuses. J Biol Chem 226: 497–509.
10. Parekh AC, Jung DH (1970) Cholesterol determination with ferric choloride – uranium
acetate and sulphuric acid-ferrous sulphate reagents. Anal Chem 42: 1423–1427.
11. Rice EW (1970) Triglycerides in serum. In Rodernick D and Donald (eds), Standard
Methods of Clinical Chemistry: Academic press, New York, pp 215–222.
12. Zilversmit BB, Davis AK (1957) Micro determination of plasma phospholipids by
trichloro acetic acid precipitation. J Lab Clin Med 35: 155–160.
13. Lopes-Virella MF, Stone P, Ellis D, Colwell JA (1977) Cholesterol determination in HDL
separated by three different methods. Clin Chem 23: 882–885.
14. Reitman S, Frankel S (1957) A colorimetric method for determination of serum glutamic
oxaloacetic and glutamic pyruvic transaminase. Am J Clin Pathol 28: 56–63.
15. King J (1965) In Practical clinical enzymology. D Van Nostrand Co Ltd, Landon, pp
83–93.
16. Lowry OH, Rose Brough NJ, Randal RJ (1951) Protein measurments with the folin
phenol reagent, J Biol Chem 193: 265–275.
17. Saleena Mathew, Menon PVG, Kurup PA (1986) Effect of administration of carnitine on
the severity of myocardial infarction induced by isoproterenol in rats. Aust J Exp Biol
Med Sci 64: 79–87.
18. Wexler BC, Greeberg BP (1978) Protective effects of clofibrate on isoproterenol induced
myocardial infarction in atherosclerotic and non atherosclerotic rats. Atherosclerosis 29:
373–395.
19. Rathore N, Kahe M, John S, Bhatnagar D (2000) Lipid peroxidation and antioxidant
enzymes in isoproterenol induced oxidative stress in rat erythrocytes. Ind J Phy Phar 44:
161–166.
20. Manjula TS, Devi CS (1993) Effect of aspirin in mitochondrial lipids in experimental
myocardial infarction in rats. Biochem Mol Biol Int 29: 921–928.
21. Saleena Mathesw, Menon PVG, Kurup PA (1981) Changes in myocardial and aortic
lipids, lipolytic activity and faecal excretion of sterols and bile acids in isoproterenol
induced myocardial infarction in rats. Ind J Biochem Biophys 18: 131–133.
22. Msengi AE, Mbise RL, Msuya PM, Amsi DM (1985). The biochemistry of water from
unripe coconuts obtained from two localities in Tanzania. E Aft Med J 62: 725.
23. Chavalittamrong B, Pidatcha P, Thavisri V (1982) Electrolytes, sugar, calories, osmolar-
ity and pH of beverages and coconut water. Southeast Asian J Trop Med Pub Health 13:
427–431.
24. Suter PM (1999) Effects of K, Mg, Ca and fiber on risk of stroke. Nut Rev 57: 84–88.
25. Syme SI, Marmot MG, Kagan A, Kato H, Rhoads G (1975) Epidemological studies of
coronary heart disease and stroke. Am J Epid 102: 477–480.
26. Naik P, Malati T, Ratnakar KS, Naidu MUR, Rajasekhar A (1999) Cardio protective
effect of magnesium chloride in experimental acute myocardial infarction. In J Exp Biol
37: 131–137.
27. Raghu C, Peddeswara Rao P, Seshagiri Rao D (1999) Protective effect of intravenous
magnesium in acute myocardial infarction following thrombolytic therapy. Int J Cardiol
71: 209–215.
28. Rubenowitz E, Molin I, Axelsson G, Rylander R (2000) Magnesium in drinking water
in relation to morbidity and mortality from acute myocardial infarction. Epidemiology
11: 416–421.
29. Gyamlani G, Parikh C, Kulkarni AG (2000) Benefits of magnesium in acute myocardial
infarction: timing in crucial. Am Heart J 139: 703–707.
30. Shechter M, Merz CN, Paul-Labrador M, Meisel SR, Rude RK, Molly MD, Dwyer JH,
Shah PK, Kaul S (1999) Oral magnesium supplementation inhibits platelet-dependent
thrombosis in patients with coronary artery disease. Am J Cardiol 89: 152–156.
31. Padmakumar Nair KG (1997) Biochemical studies on coconut protein. Ph.D thesis,
University of Kerala, Thiruvananthapuram, India.
32. Raiver HB, Stefamie MBB, Ralf PB, Laddaval P, Michael P, Reinhold N, Andrias M,
Jurgen CF (1997) Dietary L-arginine reduces the progression of atherosclerosis in
cholesterol fed rabbits-comparison with lovartatin. Circulation 96: 1282–1290.
33. Jayalekeshmy A, Arumugan C, Narayanan CS, Mathew AG (1986) Changes in chemical
composition of coconut water during maturation. J Food Sci Tech 23: 409–412.