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_________________________________* Corresponding author:S.Selvadurai,Department of Pharmacognosy, Smt.Sarojini Ramulamma College of Pharmacy, Mahbubnagar, A.P.E-mail address: sasdurai85@gmail.com

Available Online at: www.ijrpp.com Print ISSN: 2278 - 2648 Online ISSN: 2278 - 2656

(Research article)

ANTI-DIABETIC ACTIVITY OF WHOLE PLANT OF SIDA SPINOSALINN. (MALVACEAE) ON DIABETIC INDUCED RATS

*1S.Selvadurai, 2R.Senthamarai, 2T.Shri Vijaya Kirubha, 3G.Nagarajan, 3Gayasuddin Mouid Md.1Department of Pharmacognosy, Smt.Sarojini Ramulamma College of Pharmacy, Mahbubnagar, A.P.2Department of Pharmacognosy, Periyar College of Pharmaceutical Sciences, Trichy – 600021, T.N.3Department of Chemistry, Smt.Sarojini Ramulamma College of Pharmacy, Mahbubnagar, AP.

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ABSTRACT In the present study, the anti diabetic effect of ethanolic extract of Sida spinosa Linn. (Malvaceae) whole plant was studied in normal and alloxan induced (120 mg/kg, Single intraperitoneal injection) diabetic rats. Anti-Diabetic activity in normal rats was tested after administration of 200 mg/kg and 400 mg/kg of test extract. Glibenclamide was used as reference drug and showed significant anti-diabetic effects in normal rats. In anti-diabetic study a dose of 200 mg/kg and 400 mg/kg of ethanolic extract has shown reduction in triglycerides (TG), Cholesterol and Glucose level. The results obtained from the experiment provided scientific evidence in favour of the traditional use of Sida spinosa Linn. whole plant for the treatment of diabetes mellitus.Key Words: Anti-Diabetic, Sida spinosa Linn., Alloxan, Glibenclamide.

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INTRODUCTIONDiabetes is one of the most challenging diseases facing health care professionals today. Its increasing prevalence puts a large burden on society and the public health sector [1]. Type 1 diabetes is characterized by an absolute deficiency of insulin secretion, associated with auto-immune destruction of pancreatic β-cells, and this disease is more likely to occur in relatives of an affected person [2]. Type 2 diabetes, which accounts for more than 90% of cases, is caused by a combination of resistance to insulin action and impaired insulin secretion [3]. Plants have been reported as an exemplary source of drugs, and many of the currently available drugs have been derived directly or indirectly from them. In recent years, there has been renewed interest in plant medicine [4-6]. For the treatment against

different diseases as herbal drugs are generally out of toxic effect [7, 8]. Reported from research work conducted on experimental model animal. Although in human, whether there is any toxic effect are not investigated. It is reported that about 800 plants may possess anti-diabetic potential[9].Isolated studies screened various plants having “folk medicine reputation” by biochemical test for this Anti-Diabetic effect [10]. The plant Sida spinosa Linn. has been claimed to possess various medicinal properties. The root, leaves and fruits destroy “Kapha” and “Vata” tonic in wasting diseases, cure ulcers and biliousness, useful in urinary discharges, scalding urine, leprosy, and skin infections. The fruit is also astringent and cooling [11, 12]. Sida spinosa Linn. is used in the treatment of asthma and other chest ailments and as a tonic [13]. The leaves have

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reportedly been used for treatment of some skin diseases and as oral snake bite treatment [14]. The roots and leaves of Sida spinosa Linn. are used in treatment of diarrhoea and dysentery [15]. The leaves are demulcent and refrigerant, and are useful in cases of gonorrhoea, gleet and scalding urine [16].The decoction of the root-bark and root is used as a demulcent in irritability of the bladder and in gonorrhoea [17,18]. The root acts as a gentle tonic and diaphoretic, and is complied in mild cases of debility and fever[19]

The present investigation is concerned with the widely distributed indigenous medicinal plant Sida spinosa Linn. From the survey and relevant literature of the plant Sida spinosa Linn. it is most well known in traditional medicine practices and less work has been performed on different parts of the plant. The study investigated the Anti-Diabetic effects of Sida spinosa Linn. Whole plant in normal, and alloxan induced diabetic rats.

MATERIALS AND METHODSCollection of plantThe plant specimen for the proposed study were collected from Komaneri village, Thothukudi Dist. Tamilnadu in the month of January 2011, the plant material was identified and authenticated by Dr. P. Jayaraman, Plant Anatomy Research Centre, Pharmacognosy Institute, Chennai. Voucher no. PARC/2011/825.

Extraction of plant materialThe plant material was dried in the shade, then the shade dried plant material was subjected to get coarse powder and it was extracted in soxhlet apparatus using various solvents according to their polarity. The marc left after acetone extraction was dried and extracted with 2-3 litres of ethanol 95%

by continuous hot percolation using soxhlet apparatus. After completion of extraction, it was filtered and the solvent was removed by distillation under pressure. The extract was stored in desiccators [20]

Toxicity evaluation in Mice (OECD, 423 Guidance)The Ethanol extract was tested for its acute toxicity in mice. To determine acute toxicity of a single oral administration of the herbal drug, different doses of the drug (10, 100, 500, 1000, and 2000 mg/kg) were administrated to different groups of mice (3 mice were used for each group, control mice received Normal saline). Mortality and general behaviour of the animals were observed periodically for 48 hrs. The animals were observed continuously for the initial 4 hrs and intermittently for the next 6 h and then again at 24 hrs and 48 hrs following drug administration.

Effect of Alloxan - Induced diabetic ratsMale Wistar Albino Rats (180 – 200g) were made diabetic by single IP injection of 120 mg/kg body weight of alloxan monohydrate (Spectrochem Pvt.ltd. Mumbai-India) (5% w/v in water). After 4 days blood samples were withdrawn and glucose levels were determined to confirm the development of diabetes (>350 mg/100 ml)

Experimental DesignThe Anti-Diabetic activity of ethanol extract of Sida spinosa Linn. were studied by using Albino Rats. The diabetes was induced chemically using Alloxan. Male Wistar Albino Rats (180 – 200g) were used for the study. Animals were fasted for 16 hours but were allowed for free access to water prior to the experiment. Fasted rats were divided into five groups of six animals each.

Group I - - Served as Control (0.9%W/V Saline)Group II - Diabetic control (Alloxan 120 mg/kg)Group III - Standard drug (Glibenclamide 10 mg/kg)Group IV - Ethanol Extract of Sida spinosa (200 mg/kg/p.o)Group V - Ethanol Extract of Sida spinosa (400 mg/kg/p.o)

Statistical AnalysisStatistical Analysis was performed using one way analysis of variance (ANOVA) followed by

Dunnett-t Test were expressed as Mean ± SEM from six rats in each group. P- values <0.05 were considered significant.

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Table – 1 Effect of Ethanolic extract of Sida spinosa Linn. on Blood Glucose Level in alloxan induced diabetic rats

Group Treatment Blood Glucose level (BGL) in mg dL-1

Initial 4th

Day 7th

day 10th

Day

Control 85.67±1.585 87.00±1.693 86.33±2.155 83.00±4.405Diabetic Control 298.8±5.205 322.0±5.568 358.3±3.774 354.0±8.359Standard 305.8±4.929* 222.0±6.923* 175.7±4.522* 137.7±4.849*

(Glibenclamide 10 mg/kg)Ethanolic Extract 304.0±6.512* 229.3±5.766* 190.5±5.058* 190.7±2.813*

(200 mg/kg)Ethanolic Extract 306.3±6.249* 224.0±4.412* 200.3±5.920* 182.8±8.304*

(400 mg/kg)

Values are Mean ± SEM; n = 6; *P < 0.05 vs. Diabetic Control

Table – 2 Effect of Ethanolic extract of Sida spinosa Linn. on Biochemical Parameters in alloxan induced diabetic rats

S.No Parameters Control Diabetic control

Standard Glibenclamide 10 mg/kg

Ethanolic extract 200 mg/kg

Ethanolic extract 200 mg/kg

1

2

Triglycerides

Total cholesterol

44.42 ± 0.595*

108.50 ±4.332*

50.60 ± 1.569*

118.00 ±6.361*

45.43 ± 0.869*

105.6 ± 2.932*

47.60±0.757*

116.6±4.737*

46.08±1.511*

113.5±5.506*

Values are Mean ± SEM; n = 6; *P < 0.05 vs. Diabetic Control

Fig – 1 Effect of Ethanolic extract of Sida spinosa Linn. on Blood Glucose Level in alloxan induced diabetic rats

\

Blood Glucose Level (BGL) in mg dL-1

Initi

al

4th d

ay

7th d

ay

10th

day

0

100

200

300

400Diabetic Control

Glibenclamide

Ethanolic Extract 200mg/kg

Ethanolic Extract 400mg/kg

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Fig: 2 Effect of Ethanolic extract of Sida spinosa Linn. on Biochemical Parameters in Alloxan induced diabetic rats

triglycerides

Control

Diabetic

Contro

l

Gliben

clam

ide

Ethan

olic E

xtra

ct 2

00m

g/kg

Ethan

olic E

xtra

ct 4

00m

g/kg

0

20

40

60Total Cholesterol

Con

trol

Diabe

tic C

ontrol

Glib

enclam

ide

Ethan

olic Extract 200

mg/kg

Ethan

olic Extract 400

mg/kg

0

50

100

150

Histopathological StudiesA portion of pancreas tissue in each group was fixed in 10 % Formaldehyde (Formalin diluted to 10 % with normal saline) and preceded for histopathology. After paraffin embedding and

block making, serial sections of 5 µ thickness were made, stained with hematoxylin and eosin and examined under microscope. A few photomicrograph of representative type were also taken.

Fig – 3 Histopathological details of Ethanolic extract of Sida spinosa Linn.

Normal Pancreas Alloxan induced (120 mg/kg)

Standard Drug Ethanolic extract (Glibenclamide 10 mg/kg) Sida spinosa (200 mg/kg)

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Ethanolic Extract of Sida spinosa (400 mg/kg)

RESULTS AND DISCUSSIONThe plant extract of Sida spinosa Linn. showed Anti-Diabetic activity by reducing blood glucose level significantly. It is also much effective when compare with the standard drug Glibenclamide. It reduce blood glucose level up to 224.0, 200.3, 182.8 mg dL-1 at successive days of 4, 7, 10 at the dose of 400 mg/kg in rats compare with the standard drug which reduce blood glucose level upto 222.0, 175.7, 137.7 mg dL-1. We found that the ethanolic extract of plant Sida spinosa Linn. is effective in reducing blood glucose level compare to the standard drug (Glibenclamide).The cellular integrating and architecture were intact in the control group. Pancreatic section stained with hematoxylin and eosin (H&E) showed that alloxan caused severe necrotic changes of pancreatic islets, especially in the centre of islets. Nuclear changes, karyolitic, disappearance of nucleus and in some places residue of destroyed cells were visible. Relative reduction of size andnumber of islets especially around the central vessel and severe reduction of beta cells were clearly seen Diabetic control.

These was also a relative increase of granulated and normal beta cells is the diabetic group which consumed 200 mg/kg body weight ethanolic extract, when compared with the diabetic group which consumed 400 mg/kg body weight ethanolic extract. Pancreas of the diabetic group which consumed Glibenclamide 10 mg/kg body weight showed close similarity to group which consumed with test extract.

CONCLUSIONThe findings of this study indicate that consumption of the ethanolic extract of Sida spinosa Linn. exerts significant Anti-Diabetic effect in diabetic rats. Histopathological studies of the pancreas of diabetic treated rat show evidence of signs of regeneration of ß cells in groups receiving Sida spinosa Linn. in view of the restorative (protective) effects of the extract on pancreatic islet cells. Further investigation with longer period of higher doses may show clearer features of these findings.

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