Anti Diabetic Effect Of Momordica Charantia Biology Essay

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The alcohol extract of whole fruit was tested for its efficacy in Alloxan induced diabetic rabbit. The diabetic rabbits were divided into 5groups. GroupI received 2%gumacasia, groupII(positive control) received standard drug Metformin(62.5mg+2%GA), group III,IV,V(T1 T2 T3) were treated orally with daily dose of 0.5grm,1gm,1.5gm respectively for 35 days. for all diabetic rabbits after giving TEST,NC,PC preparations, the blood samples were collected and determined the blood glucose level 0,1,3,24hrs intervals. 0hr reading is before drug giving and remaining 3 readings after drugs giving. 24th hr reading is considered as 0hr reading for next day.

RESULTS: administration of alcohol of extract of bitter melon produced a dose dependent decrease in blood glucose levels in Alloxan induced rabbits. There was significant fall in blood sugar level in High dose(1.5gm/kg) in comparison to low dose(0.5gm/kg) and median dose(1gm/kg) shown by LSD test. this is comparable to the effect of Metformin.

CONCLUSION: the results of this study show that chronic oral administration of extract of Momordica charantia fruit at an appropriate dosage may be good alternative anti diabetic agent.

KEY WORDS : hyperglycemia, Metformin, Alloxan, Momordica charantia

INTRODUCTION:

Diabetes mellitus is the world largest endocrine disease with deranged carbohydrate, fat and protein metabolism. The diabetes mellitus is mainly classified into two major groups, Type-1 (insulin dependent diabetes mellitus), Type-2 (non-insulin dependent diabetes mellitus. As per WHO report, approximately 150 million people have Diabetes mellitus worldwide, and this number may well doubled by the year 2025. Statistical projection suggests that the number of diabetics will rise from 15 million in the year 1995 to 57 million in 2025 in India. This number is making India the country with the highest number of diabetics in the world (1). Long-term complications of diabetes are micro vascular (neuropathy, retinopathy, nephropathy) and macro vascular (heart complications)(2) diseases. The anti diabetic drugs are mainly used for to replace the insulin deficiency or to enhance the action of insulin and/or decrease the insulin resistance. Although many drugs and interventions are available to manage diabetes, these are expensive for the large diabetic population of developing countries like India, apart from their inherent adverse effects.(3) So it is necessary to look for new cheep alternatives to manage this major health problem. Different indigenous drugs are used in this subcontinent for several centuries for treatment of Diabetes mellitus with conflicting report of their efficacy because of lack of scientific investigation in a laboratory setting. One such plant, Momordica charantia (Karela) whose fruit is long been used traditionally in the treatment of Diabetes mellitus(6) in South Asian countries and has rich Ayurvedic reference to is selected for the study. In this study, the anti diabetic potential of this unripe fruit extract of Momordica charantia (Karela) was screened on laboratory animal model(4,5).

MATERIALS AND METHODS

Plant material: Fresh green fruits of bitter gourd popularly known as karela was obtained in sufficient quantity from local market in Nandyal, A.P. in nov 2012. They were carefully washed to remove dust particles and other foreign materials and dried in shaded area. The completely dried fruits was powdered with electric grinder and stored in well closed bottles

ALCOHOL EXTRACT PREPARATION: The extract is a concentrated preparation of vegetables or animal source Extract: The extract is the process or act of pulling or drawing out the active principle of a particular material like plants or animal organs. In the present study the percolation method was selected to extract the active principle of bitter gourd plant material. Cold percolation method: This is a traditional method of extraction used by the herbalists throughout the world. This is the original extraction method, and it continuous to be the backbone of the present extracting technology. The distillation devices are "modified Soxhlet extractions" made by Eden Labs(8) Extraction procedure: The dried fine powder of the bitter gourd was weighed on balance 25g. and taken into the sac like cloth material and placed in the Soxhlet basket. 250ml of ethyl alcohol was taken as solvent into the Soxhlet flask. Cold tap water must be flow through the inlet and outlet of the condenser. The Soxhlet apparatus keep in running for 24hours for proper extraction. The extract laden solvent falling from the Soxhlet basket is dark in color and it becomes clearer, that indicates the extraction process is finished(,9). At the end of the extraction process the solvent is then evaporated and the remaining mass is measured. The percentage yields are calculated as mg per gm dried powder. in 250ml of alcohol,25gms powder was suspended. 5gms (20%) of extract was obtained. The extract was suspended in 5ml of 2% Gum acacia and used for the oral administration in diabetic rabbits.

Animals used: 25 Rabbits of either sex, adult, healthy albino rabbits of local strain weighing between 1 to 4 kg were used in this experiment. All the animals were kept in an air-conditioned animal house of the Pharmacology department at Santhiram Medical College, Nandyal, Kurnool dist. AP. The animals, rabbits were offered a natural food like grass and leaves and allowed a tap water to drink.(10)

Preparation of diabetic rabbits: (Butt, T.A-1962, Akthar 1981) The 25 rabbits weighing between 1 to 4 kg were made diabetic by injecting intravenously 150mg/kg body weight of Alloxan monohydrate(11,12) Before giving Alloxan, the normal blood glucose levels of all rabbits were estimated. After 2hours of Alloxan injection the Dextrose (5gm) mixed with water fed to the all-diabetic rabbits orally to prevent hypoglycemic condition of rabbits with Alloxan (Jacob 1937.) (7)

After 72hours of Alloxan injection, the blood glucose levels of all surviving rabbits were determined by the glucose oxidase method.(Dr.Vijaya 1983.) The rabbits with blood glucose levels of 220 to 500mg/dl.were considered as diabetic and were employed for further study.(13)

Grouping of animals(rabbits)

All Alloxan diabetic rabbits were randomly divided into five groups (1-5) of five animals each.

GROUPS

ANIMALS

Drug

Remarks

I

NEGATIVE CONTROL

(2% GUM ACASIA)5ml

Placebo

II

POSITIVE CONTROL

(METFORMIN 62.5mg+2% GA)5ml

Positive

III

TEST (LOW DOSE 0.5gms)

ALCOHOL EXTRACT+2%GA 5ml

ExL

IV

MEDIUM DOSE (1gm)

ALCOHOL EXTRACT+2%GA 5ml

ExM

V

HIGH DOSE (1.5gm)

ALCOHOL EXTRACT+2%GA 5ml

ExH

For all the diabetic rabbits after giving test, negative control and positive control preparations, the blood samples were collected and determined the blood glucose 0, 1, 3, 24 hrs intervals. '0' hour reading is before drug giving.'1, 3, 24' hours reading is after drug giving. The 24th hour reading is considered as '0'hour reading for next day. After administration of drugs to the diabetic rabbits the blood was collected 1,3 and 24-hour interval daily up to 35 days and blood glucose level was determined with glucose oxidase method by using Glucometer for 15 days and then weakly for 3 weeks. The glucose oxidase method is more accurate, rapid and time saving method. It requires only small amount of blood. So this method is popularly used in India people suffering from diabetes for self-monitoring of blood glucose levels.(14)

RESULTS: In the present study, alcohol extract of unripe fruit of the Momordica charantia (Bitter guard) was assessed for its anti diabetic activity in Alloxan-induced diabetic rabbits. The results obtained were recorded

Table:1). Average Blood Glucose levels of all groups (I to V) before and after treatment up to 35days

S.No.

Group - I

Group - II

Group - III

Group - IV

Group - V

Before Alloxan

78.2 mg/dl

75 mg/dl

86 mg/dl

88 mg/dl

89 mg/dl

After Alloxan (72 Hrs.)

311 mg/dl

293 mg/dl

310 mg/dl

300 mg/dl

305 mg/dl

After Treatment

0 Hr.

1 Hr.

3 Hr.

24th Hr.

0 Hr.

1 Hr.

3 Hr.

24th Hr.

0 Hr.

1 Hr.

3 Hr.

24th Hr.

0 Hr.

1 Hr.

3 Hr.

24th Hr.

0 Hr.

1 Hr.

3 Hr.

24th Hr.

Day 1

311

308

307

304

293

283

279

290

301

307

308

302

300

292

292

291

314

319

312

304

Day2

304

294

306

293

290

276

276

291

302

304

299

296

291

290

285

285

304

278

273

260

Day3

243

284

313

303

291

271

275

271

296

301

300

297

285

288

293

288

260

263

262

279

Day 4

303

302

287

284

271

263

272

276

297

298

296

292

288

278

303

294

279

285

283

271

Day 5

284

277

255

283

276

256

259

270

292

291

296

292

294

274

270

269

271

295

298

276

Day 6

283

294

291

312

270

253

257

254

292

278

281

277

269

260

267

280

276

295

277

251

Day 7

312

287

278

271

254

238

244

251

277

274

283

264

260

275

279

266

251

244

247

233

Day 8

271

263

270

272

251

243

241

236

264

272

274

273

266

266

259

256

233

242

243

222

Day 9

272

271

278

287

236

226

227

231

273

270

268

264

256

255

254

245

222

231

240

209

Day 10

287

261

294

287

231

217

218

226

264

250

270

265

245

226

226

248

209

204

247

239

Day 11

287

276

282

274

226

211

222

218

265

241

252

245

248

223

216

213

239

163

213

185

Day 12

274

257

272

282

218

209

213

204

245

238

239

240

213

211

201

200

285

193

194

180

Day 13

282

296

291

301

204

191

191

185

240

236

225

235

200

200

206

201

180

187

191

161

Day 14

301

298

292

307

185

175

182

184

235

235

232

228

201

202

203

198

161

183

181

145

Day 15

307

298

298

308

184

173

171

177

228

224

225

224

198

204

203

200

145

158

157

154

3rd Week

289

286

299

332

158

149

153

163

227

232

228

214

169

175

145

166

147

141

146

151

4th Week

293

276

295

342

150

144

153

163

202

194

191

204

166

161

165

153

145

143

148

143

5th Week

272

278

274

276

109

112

130

124

184

194

199

192

149

151

146

137

120

127

131

118

DISCUSSION: The present study, alcohol extract of unripe fruit of the Momordica charantia (Bitter guard) was assessed for its anti diabetic activity in Alloxan-induced diabetic rabbits. The results obtained were recorded. A placebo-controlled sub-acute study was conducted on 5 groups of 5-diabetic rabbit models to show the hypoglycemic effect of 3 increasing doses (0.5 gm / kg, 1.0 gm / kg and 1.5 gm / kg body weight) of the alcohol extract of M. charantia suspended in gum acacia. The result was compared with the established anti-diabetic drug metformin in the dose of 150 mg per Kg body weight. Gum acacia was taken as the placebo in this study. The blood sugar level was recorded daily by Glucometer for 15 days and then once weekly for another 3 weeks. The decrease in the blood sugar level was recorded daily from the initial value and was shown in above table.Table:2). Mean blood sugar level of different groups:

Control

Metformin

ExL

ExM

ExH

R1

-18.14

-80.67

-55.02

-80.67

-142.74

R2

-36.81

-77.19

-41.79

-54.24

-69.29

R3

-14.45

-69.67

-90.66

-67.21

-94.03

R4

-52.09

-106.60

-4.10

-68.98

-160.09

R5

7.83

-50.78

-77.74

-67.98

-70.59

Variation

***

***

***

ns

***

Table:3). Summarization:

n

5

5

5

5

5

Total

-113.66

-384.91

-269.31

-339.09

-536.74

Mean

-22.73

-76.98

-53.86

-67.82

-107.35

SS

2083.48

1631.28

4546.87

351.32

7010.86

SD

22.82

20.19

33.72

9.37

41.87

SE

10.21

9.03

15.08

4.19

18.72

Here the blood sugar levels were highly decreased of a treatment with high dose of extract. The blood sugar levels are almost comes to the Normal levels. The high dose effect of extract is almost similar to Metformin effect after 35 days of treatment. The fall in the blood sugar level was summarized in Table No. 1 to 4 and ANOVA tests were done. There was significant variation in the decrease of blood sugar among the diabetic rabbit models in each group except in dose of 1 gm / Kg body weight.

ANOVA TABLE: 4

Source of Variation

SS

df

MS

F

P

Sig

Between Groups

19264.10

4

4816.02

6.16

P < 0.01

**

Within Groups

15623.82

20

781.19

Total

34887.91

24

Overall comparison between different groups of rabbits

The fall in the blood sugar was compared among the groups of animals with ANOVA. It was found that there was significant variation (P < 0.01) among the groups.

Multiple comparison tests were performed to find out the differences between the groups.

Comparison with Control:

The Dunnett's test was conducted between the control group and the groups that were given Metformin and the extracts in 3 increasing doses. As expected the fall in the blood sugar level was significant (P < 0.05) in the Metformin group. There was no significant difference in the fall in the blood sugar levels with ExL, ExM in comparison with the control but there was significant (P < 0.05) fall in blood sugar level in ExH group.

Comparison with Metformin

The effect of Extract in all the 3 doses in lowering blood sugar level showed no statistically significant difference with that of metformin in the doses used in this study. This result was checked by two post-ANOVA multiple comparison tests like LSD test of Fisher (accommodates a lot of Type I error) and FSD test of Scheffe (accommodates a lot of Type II error). Both the tests gave an identical result. It gave a strong hint that the Extracts of M. charantia were as efficient as metformin in lowering the blood sugar in diabetic rabbits and that was achieved in a broad range of doses ranging from 0.5 gm / Kg to 1.5 gm / Kg, so it might be much a safer alternative to the established drugs.

Comparison between different doses of the Extract

There was significant fall in blood sugar level in ExH dose in comparison to ExL and ExH dose in comparison with ExM as shown by LSD test. But such difference was not found in with Scheffe's test.

The present study, the hypoglycemic effect of M. charantia fruit extact was compared with metformin. Similar studies by Akhtar MS et al, in 1981 and Biyani MK et al (2003) the acute hypoglycemic effect was compared with sulphonylureas and concluded positive effect.

So the present study showed the hypoglycemic effect of alcoholic extract of unripe fruit of M. charantia in the dose ranging from 0.5 gm / Kg to 1.5 gm / Kg body weight of diabetic rabbits given orally. The hypoglycemic effect was comparable to that of the standard anti-diabetic drug Metformin in the dose of 62.5 mg / Kg body weight of rabbits. The broad dose range of hypoglycemic effect of M. charantia may be an interesting finding which may prove it safer in comparison to the established hypoglycemic drugs.

SUMMERY AND CONCLUSION: M. charantia or Karela was taken traditionally for control of diabetics in India and in other countries for long time. Three doses of alcoholic extract of the powder of unripe fruit of M. charantia were taken to study the hypoglycemic effect of in 5 groups of alloxan-induced diabetic in Rabbits. It was a placebo-controlled open study where blood sugar levels were recorded daily for 5 weeks. The study showed hypoglycemic effect of the extract in the oral dose range of 0.5 to 1.5 gm / kg body weight of rabbits. The hypoglycemic effect was comparable to that of established anti-diabetic drug Metormin in the dose of 62.5 mg / Kg. The broad dose range of the extract producing hypoglycemic effect in diabetic rabbit was an interesting observation, which requires further study.

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