Pharmacological Investigation of Protective Effects of Euphorbia hirta Whole Plant Extract, Against Duodenal Ulceration in Wistar Rats

 

 

Prabhat K Das1*, Sabuj Sahoo2, Ranjan Sethi1,  Praveen S Nayak1 and Shweta Nayak1

1GRY Institute of Pharmacy, Borawan, Khargone, MP-451228.

2University Department of Pharmaceutical Sciences, Utkal University-751004, Orissa.

 

 

ABSTRACT

Objective: The plant Euphorbia hirta is used in gastrointestinal disorders and used as an anti bacterial, anti fungal and having wound healing property. The present study was carried out to evaluate the effect of Euphorbia hirta whole plant (ethanolic and aqueous extract p.o.) on gastric and duodenal ulceration.

 

Materials and Methods: The study was carried out on duodenal ulceration model. The duodenal ulcers were induced by using Cysteamine hydrochloride (450 mg/kg). Ranitidine (20 mg/kg) was used as standard drug. Then the aqueous extract (300mg/kg) and ethanolic (250mg/kg) of the plant is used as the test drug, which is compared against the potent duodenal ulcer causing agent Cysteamine hydrochloride.

 

Results: Both the extracts of the plant Euphorbia hirta showed potent duodenal ulcer healing effect in Cysteamine induced duodenal ulceration. The aqueous extract of the plant Euphorbia hirta showed potent activity than ethanolic extract.

 

Conclusion: The plant Euphorbia hirta Linn. Increases healing of duodenal ulceration and prevents the development of experimentally induced duodenal ulceration in rats.

 

KEY WORDS: Duodenal ulceration, Cysteamine hydrochloride, buffered formaldehyde, paraffin, hematoxylin, eosin.

 

 

INTRODUCTION

The plant Euphorbia hirta is an important medicinal herb used to treat gastrointestinal disorders, including intestinal parasites, diarrhoea, heartburn, vomiting and amoebic dysentery. It is also regarded as an outstanding medication to treat respiratory system disorders, including asthma, bronchitis, hay fever, laryngeal spasms, emphysema, coughs and colds. It is a potent antipyretic and anti-inflammatory agent. [3] Its use in the treatment of jaundice, hypertension, oedema, anaemia and malaria, as an aphrodisiac. [4] [5] The plant material contains the secondary metabolites such as alkaloids, steroids, tannins and phenols.

 

It is also reported that the plant Euphorbia hirta possesses the inflammation and wound healing property. Duodenal ulcer is also a sort of inflammation associated with wound in duodenum. The anti ulcer study in stomach using this plant was reported using the pylorus ligation, aspirin, alcohol induced models. [6] So it is presumed that plant Euphorbia hirta may also possesses the anti ulcer activity in duodenal part also. The present study was to evaluate its effect on the development and healing of duodenal ulcers in appropriate animal model.

 

 

Figure No.1: Duodenum of control group in Cysteamine induced duodenal ulceration

 

MATERIALS AND METHODS:

Plant material:

The whole part of plant Euphorbia hirta plant was collected from young matured plant from Utkal University, Bhubaneswar, Orissa during the month of Nov-Dec and identified by the botanist of Department of Botany, Utkal University, Bhubaneswar by comparing with the voucher specimen present in the herbarium. After authentification fresh plant materials were collected in bulk, washed under running tap water to remove adhering dust, dried under shade and pulverized in a mechanical grinder. The coarse powder was used for further studies.

 

Experimental animals:

Male albino Wistar rats weighing between 180 to 250gm were used. The experimental protocol is approved by the institutional Animal Ethics committee 990/C/06/CPCSEA-Vanivihar, Bhubaneswar following C.P.C.S.E.A guideline.

 

Drugs and Chemicals:

Chemicals such as petroleum ether, chloroform, ethanol used in the study were of analytical grade and were procured from Merck specialties private limited, Mumbai, India. Cysteamine hydrochloride procured from Sigma Aldrich Pvt. Ltd. Hyderabad and Bangalore branch.

 

Extraction of plant material and preparation of test dose

About 200 gm of coarse dried powder of plant of the Euphorbia hirta was taken in the soxhlet apparatus and extracted successively [7] using the selected solvents in order (i.e. Pet. ether®Chloroform®Ethanol → Aqueous). The extraction for each solvent was carried out for 18 to 24 hours. The extract was collected by evaporating the solvents by slow heat treatment. Total 2kg of pulverized whole plant was subjected under solvent extraction to produce the required amount of test extract.

Figure no.2:  Duodenum of standard group in cysteamine induced duodenal ulceration

 

 

Figure no. 3:  Duodenum of Aqueous 300mg/kg treated group in cysteamine induced  duodenal ulceration

 

 

Acute toxicity studies:

In the acute toxicity test carried out in mice we take six doses and 10 mice in each dose of both aqueous and ethanolic extract i.e. 500, 1500, 2000, 2500, 3000, 4000 mg/kg body weight. All groups of test drug showed neither any toxic effect nor any lethal effect in the dose range of 500 to 4000 mg/kg body weight. The mortality was observed at a dose of 2500 and 3000 mg/kg in ethanolic and aqueous extract respectively which were determined as LD 50.So the one tenth of the dose of LD50 is selected as test drug for both the extracts. The dose limits were selected on the basis of previously performed oral acute toxicity studies in mice, in accordance with the OECD guidelines.[8]

 

Duodenal ulceration:

Cysteamine induced duodenal ulcers: [9-16]

The animals were treated with ranitidine (20mg/kg, p.o.), ethanolic dose (250 mg/kg) and aqueous dose (300/kg) prior to one hour before the administration of the cysteamine hydrochloride (450 mg/kg). After 36 hours fasting duodenal ulcer was induced by the administration of Cysteamine hydrochloride (450mg/kg) in rats. After 48 hours, all the animals were sacrificed and the duodenum is excised carefully. For histological evaluation, the stomach and duodenum are fixed in 10% aqueous buffered formaldehyde and paraffin-embedded sections are stained with hematoxylin and eosin. Ulcer index was measured by the following formula

 

 

Table No 1: Biochemical estimation of whole plant of Euphorbia hirta treated in Cysteamine-induced duodenal ulcers model

group

Treatment

Dose

Number

Scoring

Area (mm2)

Ulcer Index

I.

Control

2 ml/ kg

4.88±0.64

3.48±0.39

54.25±9.45

13.79

II.

Ranitidine

20mg/kg

0.76±0.16*

0.55±0.08***

9.46±2.14**

2.25

III.

Aqueous Extract

300mg/kg

1.07±0.29*

0.62±0.16**

17.29±2.65**

3.04

V.

Ethanol Extract

250mg/kg

4.02±0.51

2.48±0.46*

36.02±2.44*

7.5

* - P<0.05, ** - P<0.01, ***P<0.001

 

 

Ulcer Index = Ulcer Number + Ulcer Score + Ulcer Area × 10-1

 

Based on their intensity, the ulcers were given scores as follows:

0 = no ulcer, 1 = superficial mucosal erosion, 2 = deep ulcer usually with transmural necrosis, 3 = perforated or penetrated ulcer.

 

Statistical analysis:

Results of all the estimations done were indicated in terms of mean ± SEM. Statistical significance of data were evaluated by analysis of variance (One Way- ANOVA), followed by comparison between different groups using Dunnet multiple comparison test. P<0.05 level of significance was considered. The control group was compared with the standard group and all the experimental treatment groups like Aq300 and Eth2500 were compared with the control group.

 

Figure no.4:  Duodenum of Ethanol 250mg/kg treated  group in cysteamine induced duodenal ulceration

 

RESULTS:

Both the extracts of the plant Euphorbia hirta produced significant reduction in the ulcer index, the ulcer area and ulcer score, when compared to control. The aqueous extract i.e. 300mg/kg of the plant showed more significant (P<0.01) activity than the ethanolic extract i.e. 250mg/kg.

 

DISCUSSION:

The present study investigated the effect of the plant Euphorbia hirta on the duodenal ulceration. The plant showed significant effect on the healing of duodenal ulcers induced by Cysteamine hydrochloride.

 

Cysteamine induced duodenal ulcer in rat resembles that of duodenal ulcer in humans, histopathologically and pathophysiologically. Cysteamine hydrochloride inhibits the alkaline mucus secretion from the Brunner’s glands in the proximal duodenum and stimulates the rate of gastric acid secretion. Gastric emptying is also delayed and serum gastrin concentration is increased. Both the extracts of the plant significantly reduced ulcer score and ulcer area along with ulcer index. However the exact constituents and mechanism by which the plant Euphorbia hirta reduced gastric and duodenal ulcer formation and increased gastric ulcer cannot be explained by present data.

 

The result of the present investigation suggests that consumption of aqueous extract of the plant Euphorbia hirta is beneficial for patients suffering from peptic ulcer disease. The plant may produce both gastric antisecretory and gastric cytoprotective effect, resulting in increased healing of gastric and duodenal ulcers. As reported studies of anti bacterial and anti fungal activity on the plant, [17] [18] it also produces inhibition of the growth of Helicobacter pylori, which may be one of the main causes of healing of gastric as well as duodenal ulceration.

 

CONCLUSION:

Hence from all the results and discussion we finally come to the conclusion that all the doses of aqueous and ethanolic extracts were more or less effective towards both the gastric and duodenal ulcer. The aqueous extract of the plant Euphorbia hirta showed more significant activity than ethanolic extract.  Overall all the extract shows protective property against gastric and duodenal ulcer. Hence we come to the conclusion that the plant Euphorbia hirta has protective effect against the peptic ulcer.

 

ACKNOWLEDGEMENTS:

The authors are very much grateful to the Sigma Aldrich Pvt. Ltd., Hyderabad branch and Bangalore branch for their kind support to carry out this work in time. Thanks to Professor B.B.Barik, HOD of University Department of Pharmaceutical Sciences, Utkal University and Professor P.K.Panda, HOD of Pharmacology dept., for providing necessary lab facilities and kind co- operation to carry out this work in time. I also give my heartful thanks to Dr. Debasish Pradhan and Dr. S. Sahoo to suggest this topic and give their important suggestions and help me time to time during my project work.

 

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10.     Vogel H.G., Vogel W.H., Drug discovery and evaluation (Pharmacological Assays), Berlin: Springer Verlag; 1997. pp. 486-487.

11.     Dzan VJ, Haith LR Jr., Szabo S, Reynolds E.S., Effect of metiamide on the development of duodenal ulcers produced by cysteamine or propionitrile in rats, Clin Res. 1975, 23: pp.576A.

12.     Szabo S  Animal model: Cysteamine-induced acute and chronic duodenal ulcer in the rat. Am J Pathol, 1978, 93: pp.273–276.

13.     Szabo S, Haith LR Jr., Reynolds E.S., Pathogenesis of duodenal ulceration produced by cysteamine or propionitrile. Influence of vagotomy, sympathectomy, histamine depletion, H-2 receptor antagonists and hormones, Dig Dis Sci, 1979, 24: pp.471–477.

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Received on 25.09.2009

Accepted on 30.10.2009     

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Research J. Pharmacology and Pharmacodynamics 1(3) Nov - Dec. 2009; 150-153