ANTIHYPERLIPIDEMIC ACTIVITY OF Aegle marmelos (L) Corr., LEAF EXTRACT IN TRITON WR- 1339 INDUCED HYPERLIPIDEMIC RATS

Abstract

Aegle marmelos (L) Corr., (Rutaceae) leaf is ascribed with many therapeutic effects. The present study was undertaken to explore the antihyperlipidemic effect of aqueous, ethanol and chloroform extracts of the leaves of A.marmelos in Triton induced hyperlipidemic rats. The oral administration of the extracts, at dose of 250mg/1kg body weight in hyperlipidemic rats, dose dependently reduced the total cholesterol, triglycerides, low-density lipoproteins (LDL), very low-density lipoproteins (VLDL] and significantly increased the high density lipoproteins (HDL) level. Phytochemical screening revealed the presence of alkaloids, flavonoids, tannins, saponins and sitosterol in the A.marmelos leaves. The results obtained suggest marked antihyperlipidemic activity of the aqueous, ethanol and chloroform extracts of A.marmelos leaf. [PUBLICATION ABSTRACT]

Headnote

ABSTRACT

Aegle marmelos (L) Corr., (Rutaceae) leaf is ascribed with many therapeutic effects. The present study was undertaken to explore the antihyperlipidemic effect of aqueous, ethanol and chloroform extracts of the leaves of A.marmelos in Triton induced hyperlipidemic rats. The oral administration of the extracts, at dose of 250mg/1kg body weight in hyperlipidemic rats, dose dependently reduced the total cholesterol, triglycerides, low-density lipoproteins (LDL), very low-density lipoproteins (VLDL] and significantly increased the high density lipoproteins (HDL) level. Phytochemical screening revealed the presence of alkaloids, flavonoids, tannins, saponins and sitosterol in the A.marmelos leaves. The results obtained suggest marked antihyperlipidemic activity of the aqueous, ethanol and chloroform extracts of A.marmelos leaf.

V Keywords: Antihyperlipidemic; leaf extract; Aegle marmelos; Triton WR-1339.

INTRODUCTION Hyperlipidemic is a condition which can be found to be associated with overweight and obesity. Overweight and obesity are the main risk factors in diseases such as hypertension, non-insulin dependent diabetes mellitus, gallbladder and other types of cancer.1 It has been established that "Western diets" known for their high fat, high cholesterol, excess energy and low fiber contents, increase serum cholesterol and other lipid levels2 Which usually predisposes the individual to the aforementioned complications and diseases. It has been reported that complications and diseases associated with hyperlipidemic cause almost 12 million deaths each year all over the

world.3 Aegle mámelos (Tam. "Vilvam") is a moderate-sized tree of 20 to 30 ft height. It grows wild in forests, sacred grooves, gardens and temple gardens.4 The leaves contain four new alkaloids viz., 0-(3-3-dimethylallyl)- halfordinol, N-2-ethoxy-2-(4-methoxyphenyl)-ethylcinnamamide, N-2-methoxy-2-[4-(3',3'-dimethylallyloxy) phenyl]-ethylcinnamamide and N-2-methoxy-2-(4-methoxy phenyl)- ethylcinnamamide besides aegelenine and aegeline. Condensed tannins, phlobatannins, flavonoid glycosides, skimmianine and ? -sitosterol are also present.5·6 The leaves are bitter and used as febrifuge. Poultice made of the leaves is used for opthalmic and ulcers. Fresh leaves are used in West Bengal as a remedy for dropsy and beriberi associated with weakness of heart. The diluted leaf juice is used for catarrh. Aqueous and alcoholic extracts of the leaves are reported possess cardiotonic effect, like digitalis, on amphibian and mammalian hearts.7 The present study was undertaken to evaluate the hypolipidemic activity in triton

induced hyperlipidemic rats using aqueous, ethanol and chloroform extracts oí A

marmelos. MATERIAL AND

METHODS Plant

Material Fresh leaves of A marmelos were collected in the month of June 2011 from Villamuthur village, Perambalur district of Tamil Nadu, India. The plant was identified with the help of available Indian literature.8 The plant was verified with the help of specimen available of the Rapinat Herbarium, St Joseph's College, Tiruchirapalli, Tamil Nadu,

India. Preparation of Plant Powder and

Extracts Aqueous, ethanol and chloroform extracts of leaves were prepared following standard procedures. Fresh leaves were dried in an incubator for two days at 35°C, crushed in a mechanical grinder into fine powder. The powder (500g) was extracted sequentially with 2.5 liters of water, 2.5 litres of 70% ethanol and 2.5 liters of 60% chloroform in a soxhlet apparatus at 65°C until the powder became exhausted totally. The resulting extracts were filtered, concentrated and dried in-vacuo (yield 7.16, 7.68 and 8.25% w/w, respectively). The extracts were stored in desiccators for use in subsequent

experiments. Phytochemical

Analysis Preliminary phytochemical analysis was done following the method of

Hambone.9

Animals Wistar albino adult male rats (150-200g) randomly bred and maintained in the Animal House of Periyar Pharmacy College for Girls, Trichirappalli, Tamil Nadu were used for the study after prior scurtinization and approval from Institutional Animal Ethical committee (IAEC). Triton WR-1339 was purchased from SD Fines Chemical Ltd., Mumbai, India.

Experimental Design

Antihyperlipidemic potential of A marmelos in induced hyperlipidemie rats was evaluated as per the method described by Hirsch and Keller.10 Animals were divided into 5 groups of six rats per group. Test groups were administered aqueous, ethanol and chloratarm extracts at a dose of 250mg/kg body weight, respectively by oral route. The experimental designs were as follows:

Group I Normal (2ml/kg body weight)

Group II Triton WR- 1339 (100mg/kg body weight; ip only).

GroupIII Triton WR- 1339 (100mg) + aqueous extract of A marmelos leaves (250mg/kg body weight).

Group IV Triton WR-1339 (100mg) + ethanol extract of A marmelos leaves (250mg/kg body weight).

Group V Triton WR -1339 (100mg) + chloroform extract of A marmelos leaves (250mg/kg body weight).

After 48h of this acute experiment the blood was withdrawn by retro - orbital plexus from the animals of each group.

Measurements of Serum Lipid Profile

Total cholesterol (TC), Triglycerides (TG) and High Density Lipoprotein (HDL) were estimated using standard kits of Randox, Mumbai. Low Density Lipoprotein (LDL) and Very Low Density Lipoprotein (VLDL) were calculated by Friedewald's formula.11 VLDL Cholesterol = Triglycerides / 5 and LDL Cholesterol = Total Cholesterol - (VLDL + HDL Cholesterol).

Statistical Analysis

Data were statistically analysed using one-way ANOVA and expressed as mean ± S.E.M. followed by Dunnett's t-test using computerized graph pad instate version 3.05, Graph pad software, USA.

RESULTS AND DISCUSSION

The preliminary phytochemical screening revealed the presence of alkaloids, tannins, flavonoids, phytosterol, gums and mucilage and phenolic compounds. Triton WR - 1339 caused significant increase in serum total cholesterol, triglycerides, LDL, VLDL cholesterol and decrease HDL cholesterol level. The three extracts (Aqueous, Ethanol, Chloroform ) of A marmelos showed significant decreases in TC, TG, LDL and VLDL along with an increase in HDL level, dose dependently at the tested (250 mg / kg body weight), when administered in Triton WR- 1339 induced hyperlipidemic rats (Table 1).

Elevated plasma cholesterol and triglyceride levels are major risk factors of cardiovascular disease.12 The major risk due to hyperlipidemia is related to atherosclerosis and one of the initial events in this process is the accumulation of calls containing execs lipids within the arterial wall.13 Secondary plant metabolites such as flavonoids,saponins, beta - sitosterol, polysaccharides and tannins may be responsible for the antihyperlipidemic activity.14·15 However, contribution of hypolipidemic activity could be due to some compounds (alkaloids, flavonoids, tannins, phytosterol , gums and mucilage and phenolic compounds) present in the leaf extract of A marmelos. These compounds may respnsible for the antihyperlipidemic activity.

CONCLUSION

The preliminary study has been able to demonstrate hypolipidemic activity of leaf extract of A marmelos in Triton WR -1339 induced hyperlipidemic rats. Further scientific evaluation is needed to derive its molecular level mode of action.

References

REFERENCES

1. Ortiz Moreno A, Hernandez Navarro M D, Dorantes Alvrez L, Chamorro Cevallos G A and Hernandez Ortega M M; Comparative study of the hypolipidemic effect induced by different monounsaturated avocado oils. Proceedings VI World Avocado Congress (Acts VI Congreso Mundial del Aguacate). Vina Del Mar, Chile, 2007; 12-16.

2. Jingling C M D and Xiangrong L M D; Hypolipidemic effect of flavonoids from mulberry leaves in triton WR - 1339 induced hyperlipidemic mice. Asia Pac J Chile Nutr. 2007; 16(l):290-294.

3. Bakari A G, Onyemelukwe G C, Sani ? G, Aliya I S, Hassan S S and Aliya Τ M; Obesity, Overweight and underweight in Suburban northern Nigeria. Int J Diabetes Metab. 2007; 15:68-69.

4. Mathew Κ M; The Flora of the TamilNadu Carnatic, The Repaint Herbarium, St Joseph's College. 1983; Tiruchirapalli, India.

5. Nadkarni Κ M; Indian Materia Medica, Bombay Popular prakashan Pvt Ltd, Bombay. 1976.

6. Yoganarasimhan S Ν; Medical plant of India vol. 2 - Tamilnadu Regional Research Institute (AY.), 2000, Bangalore, India.

7. Anonymous. Wealth of India, CSIR, New Delhi, 1942.

8. Matthew Κ M; An Excursion Flora of Central TamilNadu, India, Oxford and JBH publishing co. Pvt.Ltd, New Delhi, 1995.

9. Harbone J B; Phytochemical Methods: A guide to modern techniques of plant analysis, 3rd Ed, Chapman and Hall, London, 1998.

10.Hirsh R L and Keller A; The pathogenesis of hyperlipidemia induced by means of surface- active agents. II Failure of exchange of cholesterd between the plasma and liver in rabbits given Triton WR-1339./ Exp Med. 1956; 104:1.

11.Friedewald W T, Ley R I and Stadrickason D S; Estimation of concentration at low density lipoprotein cholesterol in plasma without use of preparative ultracentrituge. Cline Chem. 1972; 18:499-502.

12.Garg A; Management of hyperlipidemic in IDDM patients. Diabetic care. 1994; 17:224-34.

13.Chisolm G M. The Oxidative modification hypothesis of atherogenesis: an overview. Free Radie Boil Med. 2001; 28:1815-1826.

14.Wang H X and Ng Τ ?; Natural products with hypoglycemic, hypertensive, hypocholesterolemic, ant atherosclerotic and antithrombtic activities, Life sic. 1999; 65(25):2663-2677.

15.Johns T, Chapman L, Amason J T, Mata R and Romeo J T; Phytochemistry of medicinal plants. Kluwer Academic publishers. 1995; 176.