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Hypoglycemic and Antihyperlipidemic Activity of Leaves and Stems of Alternanthera

Hypoglycemic and Antihyperlipidemic Activity of Leaves and Stems of Alternanthera

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05/21/2012

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 Journal of Pharmacy Research Vol.4.Issue 6. June 2011 P. Sampath Kumar et al. / Journal of Pharmacy Research 2011,4(6),1793-17951793-1795
Research ArticleISSN: 0974-6943
Available online throughwww.jpronline.info
*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
INTRODUCTION
Diabetic has been known to medical sciences longer than any other hereditarymetabolic disease. Nevertheless, the existing methods of this age-old illnessare not completely satisfactory. One area receiving particular attention todayis that of herbal folk medicines. Eventhough many new oral hypoglycemicagents are now available, there is high difficulty to choose the right medicationfor a longer period in the treatment either because of their side effects or highcost.
[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 thetreatment of simple stomach disorders, diarrhea, and dysentery and as a plasterfor diseased or wounded skin parts and against fever. The plant also usedagainst headache and the leaf sap is sniffed up the nose to treat Neuralgia. Thepaste 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]
Thepresent study was aimed to analyze the hypoglycemic effect of ethanolicextract of leaves and stems of 
 Alternanthera philoxeroides
in alloxan induceddiabetic mice.
MATERIALS AND METHODSSample Collection
The leaves and stems of 
 Alternanthera philoxeroides
were collected in andaround Kumbakonam during January 2008. They were washed with water andshade dried at room temperature for seven days. The dried materials wereground and powdered.
Extraction
The powdered leaves and stems were extracted using 99.99% ethanol by coldpercolation method. The powdered materials were soaked in ethanol (1:4) for48 hours at 25ºC and the filtrate was collected and distilled for recovering thesolvent, and then it was evaporated under reduced pressure at 50ºC
in vacuo
.
Hypoglycemic activityAnimals
Healthy Swiss Albino mice (6-8 weeks old) with equal weight were used throughout the study. They were maintained in a controlled environment of temperature(24
±
2
°
C). 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 chemicalsPvt. Ltd., Bioser. All other chemicals used were obtained from Ranbaxy ResearchLaboratories, Glaxo Laboratories, Reddy
s laboratories and Nice Pharmaceuticalcompany, India.
Hypoglycemic and antihyperlipidemic activity of leaves and stems of 
 Alternanthera philoxeroides
in alloxan induced diabetes
P. Sampath Kumar
1
*, B.Dheeba
1
,
 
S. Stalin
1
, M Maragatham
1
and M.Kannan
2
1.
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.
Induction of diabetes
Hyperglycemia was induced by a single intraperitoneal injection of freshlyprepared saline solution of Alloxan monohydrate (150 mg/kg) to over nightfasted 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 animalsGroup II– Hyperglycemia mice (alloxan 150 mg/kg)Group III–
 Alternantheran 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-treatedGroup VI –
 Alternanthera philoxeroites
stem extract 200mg/kg+ alloxan co-treatedGroup VII – Hyperglycemic mice treated with Metformin (14mg/kg)
Drug administration
Hyperglycemic mice were treated using ethanolic extract of leaves and stemsof 
 Alternanthera philoxeroides
dissolved in Tween 40, through oraladministration.
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 at5000 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 wereexpressed as mg/dl. Serum triglycerides and total cholesterol levels have beenestimated.
[4]
 
Serum enzymes aspartate aminotransferase (AST)
[5]
,
alanine transaminaseALT
[6]
, alkaline phosphatase ALP and acid phosphatase ACP
[7]
were measured.
Statistical analysis
The results of the present investigation were analysed by mean + standarddeviation. Significance between the groups was estimated by students t’ test.
[8]
P<0.01 is considered as significant values in the treated groups when comparedto control.
RESULTEffect on Plasma Glucose Level
The effect of ethanolic extract of 
 Alternanthera philoxeroides
leaves andstems on the blood glucose level of experimental animals was determined atvarious days interval such as 1, 15 and 30 (Table 1). After oral administrationof alloxan, there is a significant elevation of the plasma glucose levels by 2-3folds during experimental time periods when compared to the normal group.The administration of stems and leaves of 
 Alternanthera philoxeroides
extractin alloxan induced mice causes a significant reduction (P<0.01) in blood glucoselevel in diabetic mice at the days interval of 1, 15 and 30. There is no significantdifference 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
 
 Journal of Pharmacy Research Vol.4.Issue 6. June 2011 P. Sampath Kumar et al. / Journal of Pharmacy Research 2011,4(6),1793-17951793-1795
indicates that 200mg/kg of ethanolic extract of 
 Alternanthera philoxeroides
of leaf have a very good hypoglycemic effect on Alloxan induced mice and didnot change the blood glucose level in normal mice.
DISCUSSION
Diabetes is one of the most prevalent chronic diseases in the world for a longtime. It has been treated with several medicinal plant extracts through Folkloremedicine.
[9]
Because of the serious side effects produced by synthetichypoglycemic agents, they are not suitable for the better treatment.
[10]
Therefore, the search for more effective and safer hypoglycemic agents hascontinued to be an important area of active research. The present studyevaluates 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]
Alloxanspecifically damages the insulin secreting pancreatic beta-cell, resulting in adecrease in insulin release. Numerous studies demonstrated that a variety of plant extract effectively lowered the glucose level in alloxan induced diabeticanimals. The result indicated that both ethanolic extract of leaves and stemsof 
 Alternanthera philoxeroides
exhibited a significant antihyperglycemic effectin alloxan induced diabetic mice. Similar effects have been presented in otherresults such as aqueous extract.
[12]
and powder
[13]
of 
bonducella
. However, thepresent study suggests that ethanolic extract of leaves lowers the blood glucosein diabetic mice which may stimulate or regenerate the beta-cells to releasemore insulin than the stems of 
 Alternanthera
 
 philoxeroides
that supports ourhypothesis. 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 acceleratelipolysis, resulted in hyperlipidemia.Hypercholestremia and hypertrigly-ceridemia have been reported in alloxan induced diabetic mice.
[16,17]
and asignificant increase observed in our experiment to confirm the effect of alloxan induced damage. Under normal circumstances, insulin activatesenzymes lipoprotein lipase and hydrolyses triglycerides.
[18]
However in insulindeficient subject, it fails to activate the enzymes and causes hypertriglyce-ridemia. In the present investigation, the ethanolic extract of leaves andstems of 
 Alternanthera philoxeroides
and Metformin acted in a similar way bylowering 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 thatthe leaves have tremendous reducing power than stems. These results indicatedthat the leaves and stem of 
 Alternanthera philoxeroides
had a beneficial effecton the hyperlipidemia associated with hyperglycemia. These activities havealso 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 thenon specific damage caused by alloxan along with their diabetic inducingpotential. An increase in these enzyme activities reflects liver damage.
[21]
Inaccordance with these findings, alloxan treatment has a significant role in thealteration of liver functions by increase the activity of SGOT, SGPT, ALP andACP than those of normal value. On the other hand, treatment of diabeticmice with the ethanolic extract of leaves and stems of 
 Alternanthera philoxeroides
causes a significant reduction in the activity of these enzymes inplasma compared to the mean values of the diabetic group. The ethanolicextract of leaves and stem of 
 Alternanthera philoxeroides
may prevent hepaticinjury associated with diabetes.
[22]
CONCLUSION
This simple study confirmed that the leaves and stems of 
 Alternanthera philoxeroides
possess various phytoconstituents, which may contribute itshypoglycemic and hypolipidemic effect. In conclusion, the leaves of 
 Alternanthera philoxeroides
have potent efficiency against alloxan inducedcelluar damage than its stems.
REFERENCES
1.Dhanabal, SP, Kokate, CK, Ramanathan, M, Kumar, EP and Suresh, B. Hypoglycemic activityof 
Pterocarpus marsupium
Roxb. Phytotherapy Research. 2006; 20(1): 4-8.2.Sainty, I, Mehmood, Z and Mohammad F. Screening of some Indian Medicinal Plants for theirantimicrobial properties. Journal of Ethnopharmacology. 1998;62 (2): 183-93.3.Trinder, P. Insulin - Oral Hypoglycemic Agents and the Pharmacology of the Endocrine Pan-creas, Annual Clinical Biochemistry. 1969; 6: 24-25.4.Fossati, P and Lorenzo P. Serum Triglycerides determined colorimetically with an enzyme thatproduces hydrogen peroxide. Clinical chemistry. 1982; 28: 2077-2080.5.Bergmeyer,U. Anti-diabetes and hypoglycemic properties o
 Hemionitis arifolia
(Burm) Moorein rats. Journal of Ehtnopharmacology. 1974;106 (2): 192-7.6.Reitman, and Frankel Taskar SP. Plasma lipids and lipoproteins in treated diabetics. J.Postgr.Medical.1957; 29: 193-200.7.Nelson, T. Anti-diabetic activity of Methonol/Methylene chloride stem bark extracts of 
Terminaliasuperba
and
Canarium schwein furthii
on streptozotocin induced diabetic rats. Journal of Ethnopharmacology. 1966; 104(3): 306-9.
Table 1.Effect of ethanolic extract of Leaves and Stems of Alternanthera philoxeroides onplasma glucose level in normal and alloxan induced diabetic mice (mg/dl).
S.NOGroupsTreatment in days11530
1Normal91.5
±
2.390.5
±
5.690.1
±
1.72Alloxan induced mice (150 mg/kg)238.2
±
6.8289.3
±
5.2240.4
±
1.23
 Alternanthera philoxeroides
90.2
±
1.5*89.6
±
2.3*89.02
±
1.0*Leaf extract pre-treated (200mg/kg)4
 Alternanthera philoxeroides
90.75
±
2.3*89.2
±
3.6*91.5
±
0.6*Stem extract pre-treated (200mg/kg)5
 Alternanthera philoxeroides
225.2
±
5.8**140.3
±
4.8**117.01
±
0.4**Leaf extract (200mg/kg)+Alloxan co- treated6
 Alternanthera philoxeroides
230.25
±
2.3**260.3
±
2.8**180.3
±
1.4**Stem extract (200mg/kg)+ Alloxan co- treated7Hyperglycemic mice treated with227.27
±
2.7**148.2
±
2.4**114.2
±
1.6**Metformin (14 mg/kg)
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 diabeticmice. The results showed that the level of serum triglycerides and totalcholesterol have been increased in alloxan induced mice when compared tonormal mice. The administration of leaves and stems of 
 Alternanthera philoxeroides
(200mg/kg) and Metformin (14mg/kg) significantly decreasedserum triglycerides (P<0.01) and total cholesterol (P<0.01) when comparedwith control diabetic mice. The administration of stem and leaf extract of 
 Alternanthera philoxeroides
alone did not change serum triglycerides andtotal cholesterol levels in normal mice.
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 VsGroup I) by student’s ‘t’ test.
Table 2. Effect of ethanolic extract of leaves and stem of Alternanthera philoxeroideson serum triglycerides and total cholesterol in alloxan induced and normal mice(mg/dl).
1Normal69.3
±
0.637
±
2.82Alloxan induced mice (150 mg/kg)128.2
±
0.3962
±
0.93
 Alternanthera philoxeroides
 
Leaf extract70.4
±
0.27*39
±
0.2*pre-treated (200mg/kg)4
 Alternanthera philoxeroides
71.3
±
0.22*38
±
0.4*Stem extract pre-treated (200mg/kg)5
 Alternanthera philoxeroides
96.8
±
0.46**56.1
±
0.1**Leaf extract (200mg/kg)+Alloxan co- treated6
 Alternanthera philoxeroides
103.2
±
0.38**60.7
±
0.5**Stem extract (200mg/kg)+ Alloxan co- treated7Hyperglycemic mice treated with Metformin87.5
±
0.38**53.4
±
1.63**
Effect on hepatic enzymes
Intraperitoneal injection of alloxan to the mice causes a significant increasein 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 with200mg/kg of 
 Alternanthera philoxeroides
leaves and stem extract, there is asignificant reduction in the levels of SGOT, SGPT, ALP and ACP in serumwhen compared to the normal. Thus this result indicates that the ethanolicextract of leaves of 
 Alternanthera philoxeroides
lowers the levels of hepaticenzymes such as SGOT, SGPT, ALP and ACP when compared to the stems of 
 Alternanthera philoxeroides.
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 student’s ‘t’ test 
Table 3. Effect of ethanolic extract of leaves and stem of 
 Alternanthera philoxeroides
on the levels of SGOT, SGPT, ALP and ACP.
S.NoGroupsSGOTSGPTALPACP(IU/L)(IU/L)(KA units)(KA units)
1Normal46.5
±
0.450.0
±
1.8120.2
±
0.6632.4
±
0.262Alloxan induced mice (150 mg/kg)98.2
±
0.2186.7
±
0.536.4
±
0.2367.08
±
0.313
 Alternanthera philoxeroides
49.1
±
0.3*52.31?0.6*22.3
±
0.8*35.9
±
0.98*Leaf extract pre-treated (200mg/kg)4
 Alternanthera philoxeroides
50.3
±
O.8*52.90
±
1.0*22.0
±
0.11*38.2
±
0.69*Stem extract pre-treated (200mg/kg)5
 Alternanthera philoxeroides
Leaf extract70.4
±
0.4**68.2
±
0.148**24.5
±
0.27**46.7
±
0.38**(200mg/kg)+Alloxan co- treated6
 Alternanthera philoxeroides
Stem extract82.7
±
0.1**76.5
±
0.22**27.1
±
0.18**52.1
±
0.72**(200mg/kg)+ Alloxan co - treated7Hyperglycemic mice treated57
±
0.76**63
±
1.2**21.O
±
0.54**36.3
±
0.59**with Metformin (14 mg/kg)
Values are expressed as mean ± S.D; (n = 6). * P<0.01 Vs Alloxan induced group (GroupV,VI,and VII Vs Group II) by students ‘t’ test.
 
** P>0.01 Vs Normal mice (Group III and IV VsGroup I) by
 
students‘t’ test.
S.NoGroupsSerum TriglyceridesTotal cholestrol

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