P. Sampath Kumar et al.

/ Journal of Pharmacy Research 2011,4(6),1793-1795

Research Article ISSN: 0974-6943

Available online through www.jpronline.info

Hypoglycemic and antihyperlipidemic activity of leaves and stems of Alternanthera philoxeroides in alloxan induced diabetes
1.

P. Sampath Kumar 1*, B.Dheeba 1, S. Stalin 1, M Maragatham 1 and M.Kannan 2 Department of Chemistry and Biosciences, SASTRA University - Srinivasa Ramanujan Centre, Kumbakonam, Tamilnadu, India - 612 001. 2. P.G. Department of Microbiology, V.H.N.S.N.College, Virudhunagar-626 001, India.

Received on: 11-02-2011; Revised on: 16-03-2011; Accepted on:21-04-2011 ABSTRACT

The oral antihyperglycemic effect of the ethanolic extracts of leaves and stems of Alternanthera philoxeroides was investigated in alloxan induced diabetic mice. The administration of ethanolic extracts of leaves (200mg/kg) and stems (200mg/kg) of Alternanthera philoxeroides significantly reduced the plasma glucose level (p<0.01) at 62.5% and 53% respectively in alloxan induced diabetic mice (150mg/kg). Moreover the treatment of leaves and stems of Alternanthera philoxeroides significantly reduces the level of cholesterol, triglycerides and liver enzymes such as serum glutamic-oxaloacetic transaminase (SGOT), serum glutamic-pyruvic transaminase (SGPT), alkaline phosphatase (ALP) and acid phosphatase (ACP), which were increased in alloxan induced diabetic mice. These results concludethat the leaves of Alternanthera philoxeroides possess better hypoglycemic effects than the stems of Alternanthera philoxeroides.

Key words: Leaves and stems, Alternanthera philoxeroides, hypoglycemic activity, Alloxan.

INTRODUCTION
Diabetic has been known to medical sciences longer than any other hereditary metabolic disease. Nevertheless, the existing methods of this age-old illness are not completely satisfactory. One area receiving particular attention today is that of herbal folk medicines. Eventhough many new oral hypoglycemic agents are now available, there is high difficulty to choose the right medication for a longer period in the treatment either because of their side effects or high cost.[1] Alternanthera philoxeroides is a summer growing herb. It infests rivers, lakes, ponds and irrigation canals, as well as terrestrial habitats. It is used for the treatment of simple stomach disorders, diarrhea, and dysentery and as a plaster for diseased or wounded skin parts and against fever. The plant also used against headache and the leaf sap is sniffed up the nose to treat Neuralgia. The paste of the plant is used drawn out spines and it is also used to cure hernia. Alternanthera philoxeroides is also used to treat hepatitis, tight chest, bronchitis, asthma and other lungs troubles to stop bleeding and as a hair tonic. [2] The present study was aimed to analyze the hypoglycemic effect of ethanolic extract of leaves and stems of Alternanthera philoxeroides in alloxan induced diabetic mice. MATERIALS AND METHODS Sample Collection The leaves and stems of Alternanthera philoxeroides were collected in and around Kumbakonam during January 2008. They were washed with water and shade dried at room temperature for seven days. The dried materials were ground and powdered. Extraction The powdered leaves and stems were extracted using 99.99% ethanol by cold percolation method. The powdered materials were soaked in ethanol (1:4) for 48 hours at 25C and the filtrate was collected and distilled for recovering the solvent, and then it was evaporated under reduced pressure at 50C in vacuo. Hypoglycemic activity Animals Healthy Swiss Albino mice (6-8 weeks old) with equal weight were used through out the study. They were maintained in a controlled environment of temperature (242C). All animals were fed with standard pellet diet (Gold Mohar rat feed; M/S.Hindustan Lever Ltd., Mumbai) and water ad libitum . Chemicals Alloxan (chemical analytical grade) was purchased from SD fine chemicals Pvt. Ltd., Bioser. All other chemicals used were obtained from Ranbaxy Research Laboratories, Glaxo Laboratories, Reddy s laboratories and Nice Pharmaceutical company, India. Induction of diabetes Hyperglycemia was induced by a single intraperitoneal injection of freshly prepared saline solution of Alloxan monohydrate (150 mg/kg) to over night fasted mice. Control mice received similar volume of vehicle, normal saline (2ml/kg body weight) alone. Experimental protocol The animals were divided in to seven groups and each consists of six animals.
Group I Normal control animals Group II Hyperglycemia mice (alloxan 150 mg/kg) Group IIIAlternantheran philoxeroites leaf extract pre-treated (200mg/kg) Group IV Alternanthera philoxeroites stem extract pre-treated (200mg/kg) Group V Alternanthera philoxeroites leaf extract 200mg/kg + alloxan co-treated Group VI Alternanthera philoxeroites stem extract 200mg/kg+ alloxan co-treated Group VII Hyperglycemic mice treated with Metformin (14mg/kg)

Drug administration Hyperglycemic mice were treated using ethanolic extract of leaves and stems of Alternanthera philoxeroides dissolved in Tween 40, through oral administration. Collection of blood sample At the end of the experimental periods, experimental mice were sacrificed. Plasma and serum were separated from blood by centrifuging the sample at 5000 rpm for 10 min and stored in a refrigerator until analyzed. Biochemical Parameter Analysis Blood glucose was measured by Glucose oxidase method. [3] Glucose levels were expressed as mg/dl. Serum triglycerides and total cholesterol levels have been estimated. [4] Serum enzymes aspartate aminotransferase (AST) [5] , alanine transaminase ALT [6], alkaline phosphatase ALP and acid phosphatase ACP [7] were measured. Statistical analysis The results of the present investigation were analysed by mean + standard deviation. Significance between the groups was estimated by students t test.[8] P<0.01 is considered as significant values in the treated groups when compared to control. RESULT Effect on Plasma Glucose Level The effect of ethanolic extract of Alternanthera philoxeroides leaves and stems on the blood glucose level of experimental animals was determined at various days interval such as 1, 15 and 30 (Table 1). After oral administration of alloxan, there is a significant elevation of the plasma glucose levels by 2-3 folds during experimental time periods when compared to the normal group. The administration of stems and leaves of Alternanthera philoxeroides extract in alloxan induced mice causes a significant reduction (P<0.01) in blood glucose level in diabetic mice at the days interval of 1, 15 and 30. There is no significant difference have been found in the pre treated group of leaves and stems of Alternanthera philoxeroides extracts when compared to normal (P<0.01). The administration of Metformin alone shows a significant reduction (P<0.01) in the level of blood glucose when compared to the treated group. This result

*Corresponding author.
Dr.P.Sampath Kumar, Assistant professor, Department of Chemistry and Biosciences, SASTRA University - Srinivasa Ramanujan Centre, Kumbakonam, Tamilnadu, India - 612 001. Tel: +91 9843708269 E-mail: pskumar16@yahoo.com

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P. Sampath Kumar et al. / Journal of Pharmacy Research 2011,4(6),1793-1795

indicates that 200mg/kg of ethanolic extract of Alternanthera philoxeroides of leaf have a very good hypoglycemic effect on Alloxan induced mice and did not change the blood glucose level in normal mice.
Table 1.Effect of ethanolic extract of Leaves and Stems of Alternanthera philoxeroides on plasma glucose level in normal and alloxan induced diabetic mice (mg/dl).
S.NO Groups 1 2 3 4 5 6 7 Normal Alloxan induced mice (150 mg/kg) Alternanthera philoxeroides Leaf extract pre-treated (200mg/kg) Alternanthera philoxeroides Stem extract pre-treated (200mg/kg) Alternanthera philoxeroides Leaf extract (200mg/kg)+Alloxan co- treated Alternanthera philoxeroides Stem extract (200mg/kg)+ Alloxan co- treated Hyperglycemic mice treated with Metformin (14 mg/kg) Treatment in days 1 15 91.52.3 238.26.8 90.21.5* 90.752.3* 225.25.8** 230.252.3** 227.272.7** 90.55.6 289.35.2 89.62.3* 89.23.6* 140.34.8** 260.32.8** 148.22.4** 30 90.11.7 240.41.2 89.021.0* 91.50.6* 117.010.4** 180.31.4** 114.21.6**

DISCUSSION Diabetes is one of the most prevalent chronic diseases in the world for a long time. It has been treated with several medicinal plant extracts through Folklore medicine.[ 9 ] Because of the serious side effects produced by synthetic hypoglycemic agents, they are not suitable for the better treatment.[10] Therefore, the search for more effective and safer hypoglycemic agents has continued to be an important area of active research. The present study evaluates the antidiabetic effect of ethanolic extract of leaves and stems of Alternanthera philoxeroides in alloxan induced diabetic mice. Effect on blood glucose level The mechanism of hypoglycemic action probably involves the stimulation of insulin secretion either directly or indirectly from the pancreas. [11] Alloxan specifically damages the insulin secreting pancreatic beta-cell, resulting in a decrease in insulin release. Numerous studies demonstrated that a variety of plant extract effectively lowered the glucose level in alloxan induced diabetic animals. The result indicated that both ethanolic extract of leaves and stems of Alternanthera philoxeroides exhibited a significant antihyperglycemic effect in alloxan induced diabetic mice. Similar effects have been presented in other results such as aqueous extract.[12] and powder[13] of bonducella . However, the present study suggests that ethanolic extract of leaves lowers the blood glucose in diabetic mice which may stimulate or regenerate the beta-cells to release more insulin than the stems of Alternanthera philoxeroides that supports our hypothesis. Similarly, antihyperglycemic effect of Butea monospermous[14] and Terminala arjuna[15] have been recorded in alloxan induced diabetic mice. Effect on Triglycerides and Cholesterol In diabetic mice the utilization of impaired carbohydrate leads to accelerate lipolysis, resulted in hyperlipidemia.Hypercholestremia and hypertriglyceridemia have been reported in alloxan induced diabetic mice.[16,17] and a significant increase observed in our experiment to confirm the effect of alloxan induced damage. Under normal circumstances, insulin activates enzymes lipoprotein lipase and hydrolyses triglycerides. [18] However in insulin deficient subject, it fails to activate the enzymes and causes hypertriglyceridemia. In the present investigation, the ethanolic extract of leaves and stems of Alternanthera philoxeroides and Metformin acted in a similar way by lowering the triglycerides level by activation of lipoprotein lipase. In addition, treatment of mice with ethanolic extract of leaves and stem of Alternanthera philoxeroides markedly decrease the level of total cholesterol. Among that the leaves have tremendous reducing power than stems. These results indicated that the leaves and stem of Alternanthera philoxeroides had a beneficial effect on the hyperlipidemia associated with hyperglycemia. These activities have also been recorded with other antidiabetic plants i.e., Momordica charantia [19] and Cinnamomum tamala .[20] . Effect on Liver enzymes Serum enzymes such as SGOT, SGPT, ALP and ACP are used to evaluate the non specific damage caused by alloxan along with their diabetic inducing potential. An increase in these enzyme activities reflects liver damage.[21] In accordance with these findings, alloxan treatment has a significant role in the alteration of liver functions by increase the activity of SGOT, SGPT, ALP and ACP than those of normal value. On the other hand, treatment of diabetic mice with the ethanolic extract of leaves and stems of Alternanthera philoxeroides causes a significant reduction in the activity of these enzymes in plasma compared to the mean values of the diabetic group. The ethanolic extract of leaves and stem of Alternanthera philoxeroides may prevent hepatic injury associated with diabetes. [22] CONCLUSION This simple study confirmed that the leaves and stems of Alternanthera philoxeroides possess various phytoconstituents, which may contribute its hypoglycemic and hypolipidemic effect. In conclusion, the leaves of Alternanthera philoxeroides have potent efficiency against alloxan induced celluar damage than its stems. REFERENCES
1. 2. 3. 4. 5. 6. 7. Dhanabal, SP, Kokate, CK, Ramanathan, M, Kumar, EP and Suresh, B. Hypoglycemic activity of Pterocarpus marsupium Roxb. Phytotherapy Research. 2006; 20(1): 4-8. Sainty, I, Mehmood, Z and Mohammad F. Screening of some Indian Medicinal Plants for their antimicrobial properties. Journal of Ethnopharmacology. 1998;62 (2): 183-93. Trinder, P. Insulin - Oral Hypoglycemic Agents and the Pharmacology of the Endocrine Pancreas, Annual Clinical Biochemistry. 1969; 6: 24-25. Fossati, P and Lorenzo P. Serum Triglycerides determined colorimetically with an enzyme that produces hydrogen peroxide. Clinical chemistry. 1982; 28: 2077-2080. Bergmeyer,U. Anti-diabetes and hypoglycemic properties of Hemionitis arifolia (Burm) Moore in rats. Journal of Ehtnopharmacology. 1974;106 (2): 192-7. Reitman, and Frankel Taskar SP. Plasma lipids and lipoproteins in treated diabetics. J.Postgr. Medical.1957; 29: 193-200. Nelson, T. Anti-diabetic activity of Methonol/Methylene chloride stem bark extracts of Terminalia superba and Canarium schwein furthii on streptozotocin induced diabetic rats. Journal of Ethnopharmacology. 1966; 104(3): 306-9.

Values are expressed as mean S.D; (n = 6). * P<0.01 Vs Alloxan induced group (Group V, VI and VII Vs Group II) by students t test. ** P>0.01 Vs Normal mice (Group III and IV Vs Group I) by students t test.

Effect on plasma Triglycerides and total Cholesterol Table 2 shows that the effect of ethanolic extracts of the Alternanthera philoxeroides on serum triglycerides and total cholesterol in normal and diabetic mice. The results showed that the level of serum triglycerides and total cholesterol have been increased in alloxan induced mice when compared to normal mice. The administration of leaves and stems of Alternanthera philoxeroides (200mg/kg) and Metformin (14mg/kg) significantly decreased serum triglycerides (P<0.01) and total cholesterol (P<0.01) when compared with control diabetic mice. The administration of stem and leaf extract of Alternanthera philoxeroides alone did not change serum triglycerides and total cholesterol levels in normal mice.
Table 2. Effect of ethanolic extract of leaves and stem of Alternanthera philoxeroides on serum triglycerides and total cholesterol in alloxan induced and normal mice (mg/dl).
S.No Groups 1 2 3 4 5 6 7 Serum Triglycerides 69.3 0.6 128.20.39 70.40.27* 71.30.22* 96.80.46** 103.20.38** 87.50.38** Total cholestrol 372.8 620.9 390.2* 380.4* 56.10.1** 60.70.5** 53.41.63** Normal Alloxan induced mice (150 mg/kg) Alternanthera philoxeroides Leaf extract pre-treated (200mg/kg) Alternanthera philoxeroides Stem extract pre-treated (200mg/kg) Alternanthera philoxeroides Leaf extract (200mg/kg)+Alloxan co- treated Alternanthera philoxeroides Stem extract (200mg/kg)+ Alloxan co- treated Hyperglycemic mice treated with Metformin

Values are expressed as mean S.D; (n = 6). * P<0.01 Vs Alloxan induced group (Group V, VI and VII Vs Group II) by students t test. ** P>0.01 Vs Normal mice (Group III and IV Vs Group I) by students t test

Effect on hepatic enzymes Intraperitoneal injection of alloxan to the mice causes a significant increase in the activities of SGOT, SGPT, ALP and ACP to 98.2 0.21, 86.7 0.5, 36.4 0.23 and 67.08 0.31 respectively (Table 3). After the treatment with 200mg/kg of Alternanthera philoxeroides leaves and stem extract, there is a significant reduction in the levels of SGOT, SGPT, ALP and ACP in serum when compared to the normal. Thus this result indicates that the ethanolic extract of leaves of Alternanthera philoxeroides lowers the levels of hepatic enzymes such as SGOT, SGPT, ALP and ACP when compared to the stems of Alternanthera philoxeroides. Table 3. Effect of ethanolic extract of leaves and stem of Alternanthera philoxeroides on the levels of SGOT, SGPT, ALP and ACP.
S.No Groups 1 2 3 4 5 6 7 Normal Alloxan induced mice (150 mg/kg) Alternanthera philoxeroides Leaf extract pre-treated (200mg/kg) Alternanthera philoxeroides Stem extract pre-treated (200mg/kg) Alternanthera philoxeroides Leaf extract (200mg/kg)+Alloxan co- treated Alternanthera philoxeroides Stem extract (200mg/kg)+ Alloxan co - treated Hyperglycemic mice treated with Metformin (14 mg/kg) SGOT (IU/L) 46.50.4 98.20.21 49.10.3* 50.3O.8* 70.40.4** 82.70.1** 570.76** SGPT (IU/L) 50.01.81 86.70.5 52.31?0.6* 52.901.0* ALP (KA units) 20.20.66 36.40.23 22.30.8* 22.00.11* ACP (KA units) 32.40.26 67.080.31 35.90.98* 38.20.69* 46.70.38** 52.10.72**

68.20.148** 24.50.27** 76.50.22** 631.2** 27.10.18**

21.O0.54** 36.30.59**

Values are expressed as mean S.D; (n = 6). * P<0.01 Vs Alloxan induced group (Group V,VI,and VII Vs Group II) by students t test. ** P>0.01 Vs Normal mice (Group III and IV Vs Group I) by studentst test.

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8. 9. 10. 11. 12. 13. 14. Snedeeor, GW, Cochran, WG. Clinical chemistry laboratory manual Statistical Methods. Najih. A. Naser Saleh, A. Naser. Oxford and IBH Publishing Co., New Delhi, 1967. pp 137-145. Akhtar, FM and Ali, MR. Study of the antidiabetic effect of a compound medicinal plant prescription in normal and diabetic rabbit. Journal of Pak Med Assoc. 1984.34-239. Larner, J. The pharmacological basis of therapeutics. 7th Ed. New York. Macmillan. Journal of Public Health Med. 1985; 16: 263-269. Carson, J.F. Chemistry and biological properties of onion and garlic. Food Rev. Interrnet. 1987; 3: 71-103. Simon, A, Najid, A, and Chulio, A. Traditional plant medicines as treatment for diabetes. Diabetes care. 1987; 12: 553-64. Rao, N, Dwiredi, S.K, and Swarup, D. Hypoglycemic and antihyperglycemic effects of Alternanthera philoxeroides leaves in rabbits. Current science. 1994; 69: 932-933. Miura, T, Kubo M, Itoh Y, Iwamoto N, Kato M, Park S.R, Ukawa Y, Kita Y, Suzuki I,. Antidiabetic Activity of Lyophyllum decastes in Genetically Type 2 Diabetic Mice. Biology of pharmacological bulletin. 2002; 5: 1234-1237. Ragavan, B and Krishnakumari, S. Antidiabetic effect of terminala arjura bark extract in alloxan induced diabetic rats. Indian Journal of clinical Biochemistry. 2006; Vol 21 (2); 123-128. 16. 17. 18. Pari, L. Antihyperlipidemic effect of Scoparia dulcis (Sweet btoom weed) in Streptozotocin diabetic rats. Journal of Medicinal food. 2004; 9(1): 102-7. Sharma, SR, Dwivedi, SK and Swarup, D. Antihyperglycemic and insulin release effects of Aegie marmelos leaves in streptozotocin diabetic rats. Phytother. Res. 1996 10: 426-428. Taskinen, M.R.,. Lipoprotein and apoproteins in diabetes. In: Belfiore, F.,Bergnan, R.N., Molinatt, G.M. (Eds.), Current Topics in Diabetes Research,Informa Health Care. 1993; vol. 12: pp. 122 134. Singh SN, Vats P and Suri S. Effect of an antidiabetic extract of Catharanthus roseus on enzymic activities in streptozotocin induced diabetic rats J Ethnopharmacol. 2001; 76:269-77. Sharma, SR, Dwivedi, S.K., Swarup, D., 1996b. Hypoglycaemic and hypolipidemic effects of Cinnamomum tamala Nees leaves. Ind J Exp Biol. 34:37274. Hultcrantz, R, Aubert, RE and ;Herman, WH. Liver investigation in 149 asymptomatic patients with moderately elevated activies of serum aminotransferases. 1986; 21(1):109-13. Sheweita, SA, E1-Gabar, M.A.and Bastawy, M. Carbon tetrachloride induced changes in the activity of phase 11 drug metabolizing enzyme in the liver of male rats. Toxicology. 2001; 165:271-224.

19. 20. 21. 22.

15.

Source of support: Nil, Conflict of interest: None Declared

Journal of Pharmacy Research Vol.4.Issue 6. June 2011

1793-1795