INVESTIGATION OF MEDICINAL PLANTS FOR ANTI DIABETIC … fileINVESTIGATION OF MEDICINAL PLANTS FOR...

INVESTIGATION OF MEDICINAL PLANTS FOR

ANTI DIABETIC ACTIVITY

Synopsis submitted to

VINAYAKA MISSIONS UNIVERSITY

Salem

Towards Partial Fulfillment for the Award of Degree of

DOCTOR OF PHILOSOPHY

Pharmaceutical Sciences

Submitted By

D.HEPCY KALARANI, M.Pharm.,

Regd. No: D763600005

Under the guidance of

Dr.A.DINAKAR, M.Pharm.,Ph.D.,

Principal

Sun Institute of Pharmaceutical Education and Research-Nellore

2013

1

1. Introduction

Herbal drug research gained momentum in the latter part of the 20th

century with increased demand for herbal drugs in developing countries, under

developed countries. Herbal drugs constitute a diverse range of plant materials

such as herbs, roots, bark, leaves, flowers, seeds and other plant parts. In herbal

drugs, several of the constituent metabolites or at least one of the metabolites

exhibit medicinal properties. Scientific validation of existing and new herbal drugs

is extremely significant before they are put to clinical applications. Standardized

drugs provide authentic and reliable range of active principles. Scientific

validation of herbal drugs through standardization with reference to the

identification and isolation of phytochemicals is an ongoing process. The

heightened interest in standardized products is due to the belief that

standardization is directly related to the potency of the extract [1,2].

Diabetes is one of the major crippling diseases in the world leading to huge

economic losses. The persons suffering from this metabolic disease is

considered to ‘die-a-bit’ and hence ‘die-a-bit-is’ (diabetes) [3]. Gastrointestinal

(GI) disorders are common among all people, including those affected by

diabetes. As many as 75% of patients visiting diabetes clinics report significant

GI symptoms. The entire GI tract can be affected by diabetes from oral cavity

and esophagus to the large bowel and anorectal region. Ulcer disease is a

common problem in patients with or without diabetes and affects upto 10% of the

population at some time during their lives [4]. Recent research suggests that

2

inflammation inside the body plays a role in the development of tyupe 2 diabetes.

An anti-inflammatory diet and exercise plan can help prevent and treat type 2

diabetes [5].

Diabetes causes a specific form of nerve damage called neuropathy, which

cause pain in several ways [6]. Diabetes mellitus has a number of long-term

effects on genitourinary system. These effects predispose to bacterial urinary

tract infections in the patient with diabetes mellitus [7].

2. Review of literature

Literature review of selected plants [8]

Plant description of Alangium Salvifolum Wang

Family: Alangiaceae

Vernacular Names:

English: Sage leaved alangium, Ancole, Stone mango.

Tamil: Azhinjil

Telugu: Aankolam, Nalla oodaga.

3

Habitat:

It is commonly distributed in most parts of Chittoor district of Andhra

Pradesh like Tirupati, Talakona, Chandragiri and Aragonda.

Parts Used: Root bark, leaves, fruits and seeds.

Monograph: It is a deciduous shrub or tree.

Therapeutic uses:

The root bark is used for snake bite,

cutaneous troubles, anthelmintic, astringent, purgative,

diaphoretic and colic. Leaves are used in diabetes and

the fruits are used as astringent, tonic and laxative,

whereas the seeds are used in hemorrhage.

Plant description of Pavonia Zeylanica Cav

Family: Malvaceae

Vernacular Names:

Common: karubenda

Tamil: Chitta mutti

Telugu: Chittimulli, Peramuthi.

4

Habitat:

It is very commonly distributed weed in farm fields, wastelands and rare in

forest fringes, throughout the Chittoor district of Andhra Pradesh.

Parts Used: Stem, leaves and whole plant.

Therapeutic uses:

The plant is used as anti-diabetic, anti-inflammatory, laxative, Anti-

tumor, febrifuge and anthelmintic.

An extended literature review shows that an Anti-fertility activity of the stem

bark of Alangium Salvifolium Wang in wistar female rat [9], Anti-arthritic activity of

bark extracts of Alangium Salvifolium Wang [10], Antioxidant and Antimicrobial

activities of Alangium Salvifolium (L.F) Wang root [11], Wound healing property

of alcoholic extract of leaves of Alangium Salvifolium [12], Antitumor activity of

Alangium Salvifolium against Dalton’s ascetic lymphoma [13], Antibacterial and

antidiarrhoeal activity of Alangium Salvifolium Wang flowers [14] and Larvicidal

efficacy of Pavonia Zeylanica extract against Anopheles stephensi and Culex

quinquefasciatus [15] has been reported.

However, stem and leaves of the plants are not scientifically explored for

its antidiabetic activity and associated complications of diabetes like ulcer, pain,

inflammation and bacterial infections. Hence an effort has been made to screen

5

the plants for antidiabetic, anti-ulcer, analgesic, anti-inflammatory and

antibacterial activity.

3. Need for the Study

Natural products (also referred to as secondary metabolites) encompass a

wide array of truly unique chemical structures. From the largest trees to the

smallest microorganisms, all types of living creatures make natural products.

Natural products represent some of the remarkable compounds known to

mankind. The vast array of known and yet to be discovered plants, microbes, and

marine organisms that make natural products astounding. The incredible wealth

of yet to be explored microbes, plants and marine organisms means that today’s

researchers have scarcely begun to realize the true potential of natural products

for helping solve a wide range of biomedical problems.

The future of natural products is very promising. Humans also benefit

tremendously from natural products. For many years, natural products have

served as a cornerstone of the pharmaceutical industry leading to the

development of numerous lifesaving drugs that help to treat everything from

cancer to heart disease. Today, natural products are still key components of the

drug discovery process and labs.

Hence, an effort has been made to extract, isolate, characterize and

screen the selected plants, Alangium salvifolium and Pavonia Zeylanica.

6

4. Objectives

Collection and authentication of selected plants.

Extraction and Phytochemical screening.

Acute toxicity studies.

Pharmacological studies on the plant extract.

Anti-diabetic activity

Streptozotocin induced diabetic rats.

Alloxan induced diabetic rats.

The complications which are associated with diabetes are

Gastric ulcer

Pain

Inflammation

Bacterial infections

The aim and objective of the present study is to investigate the anti-diabetic

activity of the plant extracts and screening of the plant extracts on its

associated complications.

Isolation and Characterization of chemical constituent.

7

5. Methodology

The proposed plants material of fresh stem and leaves of Alangium

Salvifolium and Pavonia Zeylanica were collected from Tirupati, Chittoor district

of Andhra Pradesh, India. The species was identified and authentified by

Dr.K.Madhava Chetty, S.V.University, Tirupati. Voucher specimens were

deposited at Dept. of Pharmacognosy for further reference.

Powdered drug was extracted successively with various solvents of

increasing polarity using Soxhlet apparatus.

Preliminary phytochemical studies were done using the procedure of

Khandelwal. Alkaloids, carbohydrates, flavonoids, glycosides, tannins, phenols,

proteins, steroids, saponins and triterpenoids were qualitatively analyzed [16].

An acute toxicity was performed for the ethanolic, aqueous and Chloroform

extracts of Alangium salvifolium and Pavonia Zeylanica to determine LD50 using

different doses of the extracts [17].

Pharmacological Studies

i) Antidiabetic activity of Ethanolic, Aqueous and Chloroform extracts of

Alangium Salvifolium and Pavonia Zeylanica [EEAS, EEPZ, AEAS, AEPZ, and

CEAS & CEPZ].

Effect of EEAS & EEPZ and AEAS & AEPZ and CEAS &AEPZ on blood

glucose level in normoglycemic rats, in glucose fed hyperglycemic rats, in

8

Streptozotocin induced diabetic rats [18] and Alloxan induced diabetic rats [19]

were investigated with the doses of 400mg/kg and 800mg/kg body weight of

each plant extract. Glibenclamide was taken as a standard drug.

ii) Anti-ulcer activity

Effect of EEAS & EEPZ and AEAS & AEPZ and CEAS &AEPZ against

Pylorus ligation induced gastric ulcer and Aspirin induced gastric ulcer in rats

were evaluated. Volume of gastric juice, PH, free acidity, total acidity, ulcer index

and percentage inhibition of ulcer was calculated. The standard drug used was

Ranitidine [20].

iii) Analgesic activity

For the evaluation of analgesic activity, effect of EEAS & EEPZ and AEAS

& AEPZ and CEAS &AEPZ by acetic acid induced writhing in mice and hot plate

method in mice were studied. Aspirin used as a standard drug.

iv) Anti-inflammatory activity

The anti-inflammatory activity was determined in albino rats by

Carrageenan induced rat paw edema method. Indomethacin was the standard

drug used [21].

9

v) Antibacterial activity

The activity of the plants against gram positive and gram negative bacteria

was evaluated by Agar well diffusion method [22].

Isolation and Characterization of chemical constituent

The extracts were subjected to thin layer chromatographic technique and a

constituent from the extracts were isolated by column chromatography. The

isolated compounds were characterized by IR, NMR and Mass spectroscopic

techniques.

Statistical Analysis: By One Way ANOVA followed by Dunnett’s test using

Graph Pad Instat 3 software.

10

6. Results and Discussion

6.1 Preliminary phytochemical Studies

The preliminary phytochemical studies of extractives of Alangium

Salvifolium and Pavonia Zeylanica were given in table-1.

Table-1: Preliminary Phytochemical Screening

S.No. Phytochemical Constituents

EEAS EEPZ AEAS AEPZ CEAS CEPZ

1 Alkaloids + + + + + + 2 Carbohydrates + + + + - - 3 Flavonoids + + + - + + 4 Glycosides + + + + + + 5 Tannins/Phenols + + + + + + 6 Proteins + + + + - - 7 Steroids - - - - + + 8 Saponins + + + + - - 9 Triterpenoids + + - - + +

+: Indicate the presence of phytochemical constituents

- : Indicate the presence of phytochemical constituents

6.2 Acute toxicity study

Acute toxicity studies of the extracts are shown in table-2. Based on the

study 400mg/kg and 800mg/kg body weight of Alangium Salvifolium and Pavonia

Zeylanica were selected for the investigation of pharmacological activities.

11

Table-2: Toxicity Study

Treatment Dose (mg/kg)

No. of animals

No. of survival

No. of death

Mortality

EEAS&EEPZ AEAS&AEPZ CEAS&CEPZ

5 3 3 0 0 50 3 3 0 0 300 3 3 0 0 2000 3 3 0 0

6.3 Anti-diabetic activity

6.3.1 Effect on Blood glucose level in Normoglycemic rats

The mean blood glucose level maintained at 92.83 to 91.00mg/dl at dose

EEAS 400mg/kg and decreased from 89.17 to 83.16 mg/dl at dose EEAS

800mg/kg. For the standard drug, the same was 89.50 to 80.50 mg/dl (Table-3).

Table-3: Effect of EEAS & EEPZ and AEAS & AEPZ and CEAS & CEPZ on

Blood Glucose Level in Normoglycemic Rats

Groups Blood Glucose Level (mg/dL) 0 hour 1 hour 2 hours 3 hours 4 hours

I Normal 90.16± 1.08

88.67± 1.28

87.16± 2.40

85.00± 2.32

91.17± 2.24

II EEAS 400mg

92.83± 1.83

83.16± 1.85

84.50± 2.28

76.33± 1.89*

91.00± 2.08

III EEAS 800mg

89.17± 2.07

83.50± 2.04

79.83± 1.60*

72.50± 1.71**

83.16± 1.96*

IV EEPZ 400mg

92.00± 1.41

83.83± 1.51

82.66± 1.38

75.17± 2.53*

90.33± 1.38

V EEPZ 800mg

90.00± 1.53

84.66± 1.08

78.33± 1.41**

72.16± 2.43**

82.66± 2.03*

12

VI AEAS 400mg

89.83± 2.07

87.16± 2.17

85.00± 2.38

82.83± 1.83

92.16± 1.76

VII AEAS 800mg

90.66± 1.98

87.00± 1.53

84.00± 1.39

77.67± 1.69*

84.00± 1.46

VIII AEPZ 400mg

89.17± 1.80

87.50± 1.73

87.16± 2.27

83.16± 1.78

90.83± 2.09

IX AEPZ 800mg

92.00± 1.95

89.33± 1.80

79.33± 1.40*

75.33± 1.71**

83.00± 1.75*

X CEAS 400mg

91.67± 1.85

85.83± 1.40

85.00± 2.42

80.16± 2.04

89.00± 2.52

XI CEAS 800mg

90.00± 2.61

86.16± 1.78

83.83± 1.47

76.50± 1.76*

87.66± 1.58

XII CEPZ 400mg

91.33± 1.48

86.00± 2.67

85.66± 1.98

79.33± 1.70

87.83± 1.07

XIII CEPZ 800mg

89.83± 1.35

84.50± 1.41

79.67± 1.40*

74.83± 3.21*

83.50± 1.65

XIV

Standard

89.50± 2.45

85.50± 1.65

77.00± 1.73**

71.50± 2.04**

80.50± 2.81**

Values are expressed as Mean±SEM (n=6). *p<0.05, **p<0.01. The blood

glucose values of group II to XIV are compared with group I.

6.3.2 Effect on Blood Glucose Levels on Glucose fed Hyperglycemic Rats

(Oral Glucose Tolerance Test)

Peak increase in blood glucose concentration was observed after 1 hour

and found to reduce significantly from 81.50 to 74.50 mg/dl at dose of EEAS

800mg/kg. Standard drug group animals showed reduction from 79.50 to 71.66

mg/dl (Table-4).

13

Table-4: Effect on Blood Glucose Levels on Glucose fed Hyperglycemic

Rats

Groups

Blood Glucose Level (mg/dL) 0 hour 1 hour 2 hours 3 hours 4 hours

I

Glucose

83.83±1.92

144.83±2.52

113.50±4.14

101.17±3.52

86.00±3.39

II

EEAS 400mg

80.00±1.88

140.00±3.02

125.00±3.18

87.00±3.88*

76.67±2.39*

III

EEAS 800mg

81.50±2.23

150.00±2.34

131.16±4.56*

86.50±3.21*

74.50±1.54**

IV

EEPZ 400mg

88.17±2.56

137.17±3.26

125.67±3.63

88.66±3.13*

77.00±1.24*

V

EEPZ 800mg

90.16±3.58

151.33±3.12

131.33±4.59*

82.17±2.72**

74.16±1.25**

VI

AEAS 400mg

82.16±1.85

144.50±2.67

132.33±3.90*

92.66±3.65

84.50±2.10

VII

AEAS 800mg

84.33±1.52

150.33±2.66

135.00±4.97**

88.17±3.50*

77.50±1.41*

VIII

AEPZ 400mg

86.00±2.30

141.00±2.48

130.17±3.82*

91.16±3.64

82.16±2.24

IX

AEPZ 800mg

87.67±2.52

152.17±3.12

133.83±4.31**

86.33±3.01*

76.67±1.61*

X CEAS 400mg

82.00±2.54

134.83±4.53

129.67±3.49

100.50±3.74

85.50±2.17

XI CEAS 800mg

84.50±1.98

137.66±4.88

127.50±4.92

97.83±3.56

84.00±1.37

XII CEPZ 400mg

86.67±2.30

134.00±3.87

125.16±4.74

97.00±3.59

84.66±2.15

XIII CEPZ 800mg

88.33±3.16

140.16±3.53

127.33±5.67

86.67±2.43*

77.50±1.52*

XIV

Standard

79.50±1.41

154.83±2.57

131.33±3.05*

82.83±3.08**

71.66±2.20**

Values are expressed as Mean±SEM (n=6). * p<0.05, **p<0.01. The blood

glucose values of group II to XIV are compared with group I.

14

6.3.3 Effect on Blood glucose level in Streptozotocin Induced Diabetic Rats

The effects of the treatments with all extracts and standard drug on blood

glucose concentration in normal and diabetic rats are shown in table-5. At the

end of experiment (15th day) blood glucose level was 166.50 mg/dl and 172.33

mg/dl in the diabetic rat treated with 800mg/kg of EEPZ and EEAS respectively.

Whereas AEPZ and AEAS at dose of 800mg/kg also showed significant

reduction in blood glucose level on diabetic rats at 15th day of the study and

Chloroform extracts showed less significant activity among all other extracts

(Table-5).


Blood Glucose in STZ Induced Diabetic Rats

Groups Blood Glucose Level (mg/dL)

Initial Day 5 Day 10 Day 15

I

Normal Control

78.83±1.19

84.83±3.16

82.50±2.42

81.00±1.51

II

Diabetic Control

276.50±2.23

279.00±4.52

306.33±3.15

312.67±1.45

III

EEAS 400mg

296.17±8.95

293.50±3.91

289.83±3.67*

286.50±3.17**

IV

EEAS 800mg

270.33±6.78

262.00±6.35*

252.17±5.71**

172.33±6.27**

V

EEPZ 400mg

300.16±8.44

292.67±3.18

288.50±2.49*

233.83±5.79**

15

VI

EEPZ 800mg

274.33±7.20

261.83±3.70*

226.66±4.69**

166.50±4.61**

VII

AEAS 400mg

295.33±6.82

291.83±3.72

289.66±3.60*

286.17±3.93**

VIII

AEAS 800mg

268.00±4.47

264.16±4.73

256.00±3.80**

227.00±3.94**

IX

AEPZ 400mg

296.16±6.64

290.00±4.75

287.33±4.01*

282.33±4.80**

X AEPZ 800mg

270.50±5.23

262.66±3.99*

232.83±4.42**

196.83±4.72**

XI CEAS 400mg

280.83±3.94

282.16±3.42

300.83±3.67

302.00±3.67

XII CEAS 800mg

272.50±3.74

273.33±3.38

287.00±4.85*

286.67±5.44**

XIII CEPZ 400mg

291.50±4.45

290.33±3.53

288.50±2.48*

282.50±3.46**

XIV CEPZ 800mg

273.33±4.30

269.00±4.26

260.33±4.74**

245.83±4.46**

XV

Standard

267.66±4.89

255.16±3.09**

219.67±5.15**

150.16±5.71**

Values are expressed as Mean±SEM (n=6). *p<0.05, **p<0.01. The blood

glucose values of group III to XV are compared with group II.

6.3.4 Effect on Blood Glucose Level in Alloxan Induced Diabetic Rats

Blood glucose level in normal and experimental animals was tested on 0,

1, 3, 5 and 7 days of drug treatment. Ethanolic extract significantly decreased the

elevated blood sugar level in dose dependent manner. EEPZ at a dose of

800mg/kg decreased blood sugar level from 209.00 to 104.66 mg/dl after seven

days treatment, which was found significant and antidiabetic activity is

comparable to that of the standard drug (Table-6).

16


Blood Glucose Level in Alloxan Induced Diabetic Rats

Groups Blood Glucose Level (mg/dL) 0 Day 1st Day 3rd Day 5th Day 7th Day

I Normal Control

81.66± 1.11

81.83± 0.94

81.50± 0.62

81.67± 2.44

81.00± 1.29

II Diabetic Control

205.17± 1.72

212.00± 3.91

219.83± 4.70

236.33± 3.71

259.00± 3.36

III EEAS 400mg

205.00± 2.08

204.17± 2.86

202.16± 4.23*

174.00± 3.56**

147.83± 2.75**

IV EEAS 800mg

206.33± 2.39

203.30± 3.24

198.00± 4.74**

160.66± 4.39**

125.17± 3.32**

V EEPZ 400mg

206.16± 2.02

203.83± 3.42

199.80± 4.31*

162.16± 3.60**

130.50± 2.64**

VI EEPZ 800mg

209.00± 2.14

202.67± 3.59

190.16± 4.08**

147.00± 3.71**

104.66± 2.87**

VII AEAS 400mg

204.83± 1.83

204.00± 3.39

202.83± 4.16*

192.66± 3.27**

172.50± 3.31**

VIII AEAS 800mg

206.67± 1.80

204.83± 3.22

201.83± 3.87*

174.67± 4.03**

142.00± 3.08**

IX AEPZ 400mg

207.50± 2.20

206.00± 3.27

203.50± 3.31*

180.83± 4.36**

155.16± 2.55**

X AEPZ 800mg

206.83± 2.21

203.66± 3.40

194.66± 3.59**

164.00± 4.01**

129.83± 2.84**

XI CEAS 400mg

206.33± 1.94

210.67± 2.80

213.33± 3.15

224.83± 3.10

244.33± 3.28*

XII CEAS 800mg

207.00± 2.29

209.83± 2.18

210.50± 3.09

218.66± 4.52*

220.00± 3.50**

XIII CEPZ 400mg

207.67± 2.31

207.33± 2.49

206.83± 3.66

202.00± 3.37**

197.16± 2.82**

XIV CEPZ 800mg

208.17± 1.96

207.00± 2.83

205.16± 3.08*

192.67± 3.25**

173.50± 3.70**

XV

Standard

208.00± 2.70

198.33± 3.14*

179.67± 3.24**

130.50± 4.50**

85.83± 2.61**

17

Values are expressed as Mean±SEM (n=6). *p<0.05, **p<0.01. The blood glucose levels of group III to XV are compared with group II.

6.4 Anti-ulcer activity

6.4.1 Effect of extracts against Pylorus ligation induced gastric ulcer

The Ethanolic and aqueous extracts of the Alangium salvifolium and

Pavonia Zeylanica in the doses of 400 and 800mg/kg, chloroform extract of

Alangium Salvifolium 800mg/kg produced a reduction in the ulcer index, gastric

volume free acidity, total acidity and raised gastric PH significantly in comparison

with control group. The percentage inhibition of ulcer was 62.92% at dose of

EEAS 800mg/kg which is comparable with that of the percentage inhibition of

standard, i.e., 70.61% .

Control Standard

EEAS 400 EEAS 800 EEPZ 400 EEPZ 800

AEAS 400 AEAS 800 AEPZ 400 AEPZ 800

18

CEAS 400 CEAS 800 CEPZ 400 CEPZ 800

6.4.2 Effect of extracts against Aspirin induced gastric ulcer

The ulcer index and the calculated percentage inhibition of ulcer against

aspirin induced gastric ulcer in rats shows 69.17% at dose of EEAS 800mg/kg,

66.73% at dose of EEPZ 800mg/kg, 65.07% at dose of AEAS 800mg/kg, 52.10%

at dose of AEPZ 800mg/kg and 50.44% at dose of CEAS 800mg/kg.

6.5 Analgesic activity

6.5.1 Acetic acid induced writhing method

In acetic acid induced writhing test, at doses of 800mg/kg the EEPZ,

EEAS, AEPZ, AEAS and CEPZ inhibited the writhing responses and the number

of writhes were significantly lower than the control group and the maximum

inhibition is seen at EEPZ 800mg/kg i.e., 54.25%. The standard drug exhibited

74.84% of inhibition.

6.5.2 Hot plate method

The results of hot plate method revealed that the reaction time for mice

was significantly increased in a dose dependent manner with the reaction time

19

from 7.16 to 9.83 sec and 6.83 to 12.83 sec at dose of 400 and 800 mg/kg of

EEAS, 6.33 to 11.00 sec and 7.00 to 13.33 sec at dose of 400 and 800 mg/kg of

EEPZ, 6.16 to 9.33 sec and 7.00 to 12.67 sec at dose of 400 and 800 mg/kg of

AEAS, 6.50 to 10.83 sec and 7.33 to 12.83 sec at dose of 400 and 800 mg/kg of

AEPZ, 6.66 to 7.67 sec and 6.50 to 9.30 sec at dose of 400 and 800 mg/kg of

CEAS, 6.67 to 10.66 sec and 6.83 to 13.16 sec at dose of 400 and 800 mg/kg of

CEPZ. The standard drug exhibited the reaction time from 6.67 to 14.00 sec.

6.6 Anti-inflammatory effect on Carrageenan induced rat paw edema

The EEAS, EEPZ, AEAS, AEPZ, CEAS and CEPZ in the doses of

800mg/kg showed the percentage inhibition 63.50%, 60.32%, 60.32%, 55.55%,

34.13% and 7.93% at 24 hour. Inhibition of edema at EEAS 800mg/kg was

higher; however the standard drug exhibited the percentage inhibition of edema

as 73.81%.

6.7 Antibacterial activity

Followed by incubation, the plates were inspected to identify the zone of

inhibition. The diameter of zone of inhibition of each extracts and standard discs

of different concentrations was recorded with the help of zone measuring scale.

Zone of inhibition was 20mm, 19mm, 17mm & 18mm for high concentration of

EEAS in different organisms. Standard drug showed zone of inhibition 22mm,

26mm, 18mm & 21mm in the same organisms.

20

6.8. Isolation and Characterization of Chemical Constituent

Spectrum of isolated compound from Alangium Salvifolium

Figure 1: IR Spectrum Figure 2: Mass Spectrum

Figure 3: NMR Spectrum Probable Structure: Di-demethoxy Alanginol

Spectrum of isolated compound from Pavonia Zeylanica

Figure 4: IR Spectrum Figure 5: Mass Spectrum

N

O H

21

Figure 6: NMR Spectrum Probable Structure: Osthole dimer

7.Conclusion

Screening of anti-diabetic activity in Streptozotocin induced and Alloxan

induced diabetic rats method indicate the efficiency of Alangium salvifolium and

Pavonia Zeylanica in the maintenance of blood glucose level in diabetic induced

rats. Administration of ethanolic extracts showed a significant decrease in blood

glucose level in diabetic rats than that of aqueous and chloroform extracts.

Ethanolic extracts of the plant Alangium Salvifoilum was significantly

effective in protecting gastric mucosa against pylorus ligation induced and aspirin

induced gastric ulcer at all the dose level studied.

Many chemical agents have been reported to produce writhing but acetic

acid and phenyl benzoquinone are the two most commonly used irritants. EEPZ

at its higher doses produces a maximum percentage of inhibition. In hot plate

method a cut off period of 15 sec was observed to avoid damage to the paws.

EEPZ 800mg/kg showed the reaction time comparable with that of standard.

O

O

O

O O

O

22

Many phlogistic agents have been used for the induction of edema, among

them Carrageenan been found to be the most suitable agent and provides a

good predictive value for anti-inflammatory potential of a novel compound. EEAS

800mg/kg showed 63.50% inhibition of paw edema, which is closer to the

inhibition by a standard drug.

The antibacterial activity of Alangium salvifolium and Pavonia Zeylanica

was proved by its zone of inhibition against various gram positive and gram

negative organisms. The activity was compared with a standard drug.

Characterization of the isolated compounds by various spectroscopic

studies has been done and the probable structure was predicted.

The results of these studies confirm that the ethanolic, aqueous and

chloroform extracts of Alangium Salvifolium and Pavonia Zeylanica has an anti-

diabetic activity. Further the plant extracts are also useful for the treatment of

complications (gastric ulcer, pain, inflammation and bacterial infection) which are

associated with diabetes.

8. References

1. Sabulal B, George V. Phytochemical techniques in herbal drug research. In:

Rakesh KS, Rajesh A, editors. Herbal drugs-A wenty first century perspective.

Jaypee Brothers Medical Publishers (P) Ltd, New Delhi; 2006: p.60.

23

2. Narasimhan S, Anitha G, Ilango K, Mohan KR. Bioassay-guided isolation of

active principles from medicinally important plants. In: Rakesh KS, Rajesh A,

editors. Herbal drugs-A wenty first century perspective. Jaypee Brothers Medical

Publishers (P) Ltd, New Delhi; 2006: p.71.

3. Dixit PP, Londhe JS, Saroj SG, Devasagayam TPA. Antidiabetic and Related

Beneficial Properties of Indian Medicinal Plants. Herbal Drugs: A Twenty First

Century Prospective. Jaypee Brothers Medical Publishers (P) Ltd: New Delhi;

2006.

4. James D, Wolosin MD, Steven V, Edelman MD. Diabetes and the

Gastrointestinal Tract. Clinical diabetes, 2000; 18(4).

5. Diabetes and Inflammation. http://diabetes.webmd.com/guide/inflammation-

and-diabetes

6. Pain Management.

https://www.bd.com/us/diabetes/page.aspx?cat=7001&id=10025

7. Patterson JE, Andriole VT. Bacterial urinary tract infections in diabetes. Infect

Dis Clin North Am, 1997; 11(3):735-50.

8. Madhava Chetty K, Sivaji K, Tulasi Rao. Flowering Plants of Chittoor District

Andhra Pradeh, India,1st edition, 2008. Student Offset Printers, Tirupati.

24

9. Murugan V, Shareef H, Rama Sarma GVS, Ramanathan M, Sureh B. Anti-

fertility activity of the stem bark of Alangium Salvifolium (Linn.F) Wang in wistar

female rats. Ind J Pharmacology, 2000; 32(6):388-399.

10. Jubie S, Jawahar N, Ruby Koshy, Gowramma B, Murugan V, Suresh B. Anti-

arthritic activity of bark extracts of Alangium Salvifolium Wang. Rasayan J.Chem,

2008; 1(3):433- 436.

11. Vineet CJ, Patel NM, Dhiren PS, Paras KP, Bhavesh HJ. Antioxidant and

Antimicrobial Activities of Alangium salvifolium (L.F) Wang. Global Journal of

Pharmacology. 2010; 4 (1): 13-18.

12. Inayathulla, Karigar AA, Shariff WR, Sikarwar MS. Wound healing property of

alcoholic extract of leaves of Alangium salvifolium. J Pharmacy Research. 2010;

3(2): 267-269.

13. Venkateshwarlu R, Venu Gopal Y, Butchi Raju A, Bhanu K, Prasad.

Antitumor activity of Alangium salvifolium against Dalton’s ascetic Lymphoma..

Medicinal Chemistry & Drug Discovery. 2012; 3(2): 122-133.

14. Zahan R, Ashik Mosaddik M, Ranjan Kumar B, Mir Imam Ibne WM.

Antibacterial and antidiarrhoeal activity of Alangium salviifolium Wang flowers.

Mol & Clin. Pharmacology. 2012; 2(1): 34-43.

15. Kamaraj C, Abdul Rahuman A, Bagavan A, Abduz Zahir A, Elango G,

Kandan P et al., Larvicidal efficacy of medicinal plant extracts against Anopheles

25

stephensi and Culex quinquefasciatus (Diptera: Culicidae).. Trop Biomed. 2010;

27(2): 211–219.

16. Khandelwal KR. Practical Pharmacognosy,Techniques and Experiments,

2004. Nirali Prakashan, Pune.

17. OECD Guideline for testing of Chemicals. Test No.423. Acute Oral Toxicity-

Acute toxic class method.

18. Soon YY, Tan BKH. Evaluation of the Hypoglycemic and Anti-oxidant

activities of Morinda officinalis in Streptozotocin-induced Diabetic rats. Singapore

Med J, 2002; 43(2): 77-85.

19. Osinubi AA, Ajayi OG, Adesiyun AE. Evaluation of the Anti-diabetic effect of

aqueous extract of Tapinanthus Butungii in male Sprague-Dawley rats. Med J Isl

World Aca Sci, 2006; 16(1): 41-47.

20. Deshpande SS, Shah GB, Parmar NS. Antiulcer activity of Tephrosia

Purpurea in rats. Ind j Pharmacol, 2003; 35: 168-172.

21. Kulkarni SK. Handbook of Experimental Pharmacology, 3rd edition, 2007.

Vallabh Prakashan, Delhi.

22. Dewanjee S, Kundu M, Maiti A, Majumdar R, Majumdar A, Mandal SC. In

vitro evaluation of Antimicrobial activity of crude extract from Diospyros

peregrine, Coccinia grandis and Swietenia macrophylla. Trop J Pharm Research,

2007; 6(3): 773-778.

26

Publications

• Hepcy Kalarani D, Dinakar A, Senthilkumar N. Evaluation of Anti-diabetic

activity of ethanolic and aqueous extracts of stem and leaves of Alangium

Salvifolium and Pavonia Zeylanica. Int J Pharm Pharm Sci., 2013, 5(2):

363-366.

• Hepcy Kalarani D, Dinakar A, Senthilkumar N. Antidiabetic, Analgesic and

Anti-inflammatory activity of Aqueous extracts of Stem and Leaves of

Alangium Salvifolium and Pavonia Zeylanica. Int. J. Drug Dev & Res.,

2012, 4(4): 298-306.

• Hepcy Kalarani D, Dinakar A, Senthilkumar N. Analgesic and Anti-

inflammatory activity of ethanolic extracts of Alangium Salvifolium and

Pavonia Zeylanica . Int. J. Pharm. Sci. Rev. Res., 2012, 17(1): 77-80.

• Hepcy Kalarani D, Dinakar A, Senthilkumar N. Screening of Anti-ulcer

activity of ethanolic and aqueous extracts of Alangium Salvifolium and

Pavonia Zeylanica in rats. Int. J. Advances in Pharm. Res., 2012, 3(12):

1264-1268.

• Hepcy Kalarani D, Dinakar A, Senthilkumar N. Antidiabetic activity of

ethanolic extracts of Alangium Salvifolium and Pavonia Zeylanica in

Streptozotocin induced diabetic rats. Int. J. Pharm. Pharm. Sci., 2012, 4(5):

337-339.

INVESTIGATION OF MEDICINAL PLANTS FOR ANTI DIABETIC … fileINVESTIGATION OF MEDICINAL PLANTS FOR...

Documents

Transcript of INVESTIGATION OF MEDICINAL PLANTS FOR ANTI DIABETIC … fileINVESTIGATION OF MEDICINAL PLANTS FOR...