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Cardioprotective effect of tender coconut water in experimental myocardial

infarction

Article  in  Plant Foods for Human Nutrition · September 2003

DOI: 10.1023/B:QUAL.0000040363.64356.05

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 Plant Foods for Human Nutrition 58: 1–12, 2003.
© 2004 Kluwer Academic Publishers. Printed in the Netherlands.

Cardioprotective effect of tender coconut water in

experimental myocardial infarction

P. ANURAG and T. RAJAMOHAN∗

Department of Biochemistry, University of Kerala, Kariavattom, Trivandrum 695581,
Kerala, India; (∗ author for correspondence; )

Received 4 January 2001; accepted in revised form 19 July 2001

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.

Key words: Cholesterol, Creatine phosphokinase, Isoproterenol, Lactate dehydrogenase, Lipo-

proteins, Myocardial infarction, Tender coconut water

Introduction

Tender coconut water (TCW) is a common drink in all coconut producing

countries. It contains sugars, vitamins, minerals, proteins, amino acids and
growth promoting factors [1, 2]. Due to the presence of proteins, sugars,
vitamins, amino acids and growth hormones, coconut water is considered to
be useful for the formulation of nutritional beverages and other food products.
It is traditionally used for the treatment of diarrhea and vomiting to prevent
dehydration of the tissues. Use of TCW is recommended in cases of gastroen-
teritis and for urinary stone dissolution [3, 4]. It is a rich source of minerals
(potassium, calcium and magnesium) which are known to reduce the risk for

GSB/Prepr: QUAL 3668/DISK Pipsnr. 381082; Ordernr.: 174-JM.KLUTEX2K (qualkap:bio1fam) v.1.2

qual3668.tex; 11/04/2004; 9:03; p.1
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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.

Materials and methods

Collection of tender coconut water. Young coconuts (Cocos nucifera) (West

Coast Tall Variety) 5–6 months of age harvested from the coconut trees grown
on the University campus were used for the study. The coconuts were de-
husked, broken carefully and liquid endosperm was collected, stored in a
freezer at 0 ◦ C and used for the three day experiment.

Animals and diet. Male albino rats (Sprague-Dawley) weighing an average

of 150–160 g were divided into two groups of 24 rats each that were treated
as follows:
Group 1 – Control rats
Group 2 – Rats given TCW.
Four ml tender coconut water per 100g body weight was given to each rat
daily. Control rats received the same volume of distilled water. The animals
were housed individually in polypropylene cages and were given standard rat
chow (Sai Feeds, Bangalore, India) and distilled water ad libitum. Temperat-
ure of the animal laboratory was maintained at 25 ± 5 ◦ C with alternate 12
hour periods of light and dark. Food intake was recorded daily, body weights
of the rats were recorded weekly. The laboratory guidelines of the University
Federation of Animal Welfare, for the use and care of animals were followed
throughout the experimental period [7].
After 45 days, the rats were distributed into the following groups:
Group 1 a – Control rats
Group 1 b – Control rats with isoproterenol
Group 2 a – TCW fed rats
Group 2 b – TCW fed rats with isoproterenol.
Myocardial infarction was produced in rats in Groups 1 b and 2b, by a
subcutaneous injection of isoproterenol at a dose of 20 mg/100 g body weight,
administered twice at an interval of 24 hours [8]. Rats given isoproterenol
displayed signs of shock, tachycardia, dyspnea, rapid respiration, anuria and
prostration. Rats surviving the second injection were deprived of food for 12
hours and were sacrificed by decapitation.
Blood samples were collected and kept in a water bath at 37 ◦ C for 30
minutes and then centrifuged (2500 rpm for 15 min) for serum collection. Tis-
sues (heart and aorta) were collected in ice cold containers for the estimation

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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

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Table 1. Activity of creatine phosphokinase (CPK), serum aspartate amino transferase
(SGOT), serum alanine amino transferase (SGPT) and lactate dehydrogenase (LDH)
in rats given isoproterenol and tender coconut water

Group CPK (IU/L) SGOTa SGPTb LDHc

1a 210.67 ± 5.4 156.32 ± 4.7 73.38 ± 2.4 205.12 ± 4.3

1b 593.43 ± 8.9∗ 302.20 ± 5.4∗ 193.63 ± 3.8∗ 1294.76 ± 17.5∗
2a 215.5 ± 5.8 143.34 ± 4.3 67.79 ± 2.8 203.05 ± 7.15
2b 328.43 ± 7.5
184.95± 3.19
118.77± 3.7
976.83 ± 10.5

Results are expressed as average of values from 6 rats ± SD.
∗ Indicates that the result is significantly different from group 1a.

Indicates that the result is significantly different from group 1b.
a µ moles of oxaloacetate liberated/min/mg protein.
b µ moles of pyruvate liberated/min/mg protein.
a µ moles of NAD+ liberated/min/mg protein.

Table 2. Activity of serum aspartate amino transferase (SGOT),

serum alanine amino transferase (SGPT) and lactate dehydro-
genase (LDH) in the heart of rats given isoproterenol and tender
coconut water

Group SGOTa SGPTb LDHc

1a 431.99 ± 6.7 376.3 ± 5.5 213.5 ± 5.3

1b 637.95 ± 7.8∗ 509.62 ± 6.5∗ 655.33 ± 8.6∗
2a 423.99 ± 5.4 366.81 ± 5.2 204.95 ± 6.9
2b 495.79 ± 5.19
406.95 ± 5.3
509.47 ± 6.7

Results are expressed as average of values from 6 rats ± SD.
∗ Indicates that the result is significantly different from group 1a.

Indicates that the result is significantly different from group 1b.
a µ moles of oxaloacetate liberated/min/mg protein.
b µ moles of pyruvate liberated/min/mg protein.
a µ moles of NAD+ liberated/min/mg protein.

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

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Figure 1. Concentration of cholesterol, HDL-C and VLDL-C+LDL-C in serum of animals

given isoproterenol and TCW (mg/100 ml).
∗ 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.

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

The present study demonstrated that the administration of isoproterenol to

rats (20 mg twice) resulted in the induction of myocardial infarction. Sim-
ilar effects have been reported by others [17, 18]. Raised values of serum

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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

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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.

reducing hyperlipidemia caused by isoproterenol administration. No areas of

degeneration and infiltration were seen in the isoproterenol rats fed TCW.
Recovery from myocardial infarction was evident from the values of marker
enzymes, CPK,SGOT,SGPT and LDH, in rats fed TCW. The isoproterenol
treated rats given TCW had a greater survival rate and very little myocardial
damage compared to the rats given isoproterenol alone.
Coconut water contains many factors, which may have beneficial effects
in reducing the myocardial infarction induced by isoproterenol. A young
coconut that is 5 to 6 months old contains 300 to 500 ml of water. The
water contains proteins, fats, sugars, minerals, vitamins, free amino acids and
growth hormones [22]. The minerals include potassium (300 mg/100 ml),
calcium (30 mg/100 ml) and magnesium (10 mg/100 ml) [23].
Increased potassium intake is likely to be beneficial in patients with coro-
nory heart disease and stroke [24]. Studies in hypertensive rats have found
that high intake of potassium protects against death from stroke [25]. Also
reports indicate an inverse association of potassium intake irrespective of

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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.

hypertensive status with stroke [24]. Clinical and epidemiological evidence

suggests that a high dietary intake of potassium is associated with lower blood
pressure which is the most important risk factor for coronory heart disease
[25].
Magnesium is a cardioprotective agent that plays a key role in neurochem-
ical transmission and muscular excitability [26]. Cardio-manifestations due to
magnesium are similar to those of potassium. Magnesium inhibits both pro-
duction of endothelial derived releasing factor (nitric oxide) and contraction
of vascular smooth muscle in the pulmonary artery and the coronary arteries
[26]. It is known to reduce the secondary rise in cardiac output without a sig-
nificant alteration in cardiac function. Due to its potent vasodilator property,
magnesium is known to prevent and relieve coronary artery spasms associ-
ated with acute myocardial infarction [27]. Magnesium in drinking water is
associated with lower mortality from acute myocardial infarction [28]. Intra-
venous magnesium therapy is a safe, effective, and inexpensive modality of
treatment in acute myocardial infarction [29]. Oral magnesium supplement-

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9

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

ation inhibits platelet-dependent thrombosis in patients with coronary artery

disease [30].
TCW contains proteins such as gelatins, albumin, glutelin and prolamin. It
also contains free amino acids; arginine is the major constituent [22]. There is
evidence from earlier studies that the globulin fraction isolated from the fresh
coconut kernel has a significant hypocholesterolemic effect and the major
factor responsible for this effect is the higher arginine content [31]. Hypocho-
lesterolemic and antiatherogenic effect of L-arginine have been reported by
several investigators [6,32].
Coconut water has been reported to contain growth promoting factors
[33]. The presence of these factors is higher in the water derived from tender
coconuts. It has been reported that growth hormones improve cardiac func-
tion in experimental myocardial infarction [34]. Further studies are needed
to ascertain whether the growth hormones present in coconut water have any
beneficial effect in myocardial infarction.

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.

qual3668.tex; 11/04/2004; 9:03; p.10

 11

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.

qual3668.tex; 11/04/2004; 9:03; p.11

 12

34. Isgaard J, Kujacic V, Jennische E, Holmang A, Sun XY, Hedner T, Hjalmarson

A, Benotsson BA (1997) Growth hormone improves cardiac function in rats with
experimental myocardial infarction. Eur J Clin Invest 27: 517–525.

qual3668.tex; 11/04/2004; 9:03; p.12

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