Iranian Journal of Pharmacology and Therapeutics Razi Institute for Drug Research (RIDR) of Iran University of Medical Sciences and Health Services (IUMS) ISSN: 1735-2657 Vol. 4, Num. 1, 2005, pp. 24-27

Iranian Journal of Pharmacology & Therapeutics, Vol. 4, No. 1, 2005, pp. 24-27

Antidiarrheal Evaluation of Benincasa hispida (Thunb.) Cogn. Fruit Extracts

VRUSHABENDRA SWAMY BHYRAPUR MATHAD, SRIDHAR CHANDANAM, SREENIVASA RAO THIRUMALA SETTY, DHANAPAL RAMAIYAN, BALAMURALIDHAR VEERANNA, ASHOKA BABU VECHHAM LAKSHMINARAYANASETTRY

Department of Pharmacology (V.S.B.M.); Department of Pharmaceutical Chemistry (S.C., S.R.T.S.); Department of Pharmaceutics (D.R., B.V.); Department of Pharmacognosy (A.B.V.L.); Rural College of Pharmacy, Bangalore, India.

Address correspondence to: Vrushabendra Swamy B.M., Department of Pharmacology, Rural College of Pharmacy, Devanahalli, Bangalore–562110, Karnataka, India.E-mail: swamybm@yahoo.com

Received February 28, 2005;
Revised March 22, 2005;
Accepted April 4, 2005

Code Number: pt05006

ABSTRACT

The methanolic extract of fruit of Benincasa hispida (BHFE) was evaluated for its antidiarrheal potential against several experimental models of diarrhea in rats. BHFE treated animals showed significant inhibi-tory activity against castor oil induced diarrhea and inhibited PGE₂ induced enter pooling in rats. It also showed significant reduction in gastro intestinal motility following charcoal meal in rats. The result obtained and establishes the efficacy of BHFE as an antidiarrheal agent.

Keywords: Benicasa hispida, Antidiarrheal, Fruits, BHFE

Since the diarrhoea is leading cause of mortality in developing countries, the World Health Organization (WHO) has constituted a Diarrheal Disease Control Program (CDD), which includes studies on traditional medical practices, together with the evaluation of health education and prevention approaches [ 1 - 4 ].

The fruit of Benincasa hispida (Thunb.) Cogn., com-monly called as ash guard, belonging to cucurbitaceous is employed as a main ingredient in kusmanda lehyam, in Ayurvedic system of medicine. The leyham is used as rejuvenate agent and also numerous nervous disorders. Many empirical applications have been used in India centuries for various ailments such as GIT problems such as dyspepsia, burning sensation, heart disease, vermifuge, diabetes, and urinary disease [ 5 , 6 ]. Though some scientific studies have been carried out reveal its anti-inflammatory activity [ 7 ], diuretic activity [ 8 ] and anti cancer [ 9 ]. The major constituents of this fruits are triterpenoids, flavanoids, glycosides, saccharides, caro-tenes, vitamins, β sitosterin, and uronic acid [ 10 - 12 ]. However there is no report on antidiarrheal activity of this plant though diarrhea is common occurrence dis-ease. In the light of the above information the present investigation was undertaken to evaluate the antidiar-rheal potential of Benincasa hispida fruit extract and is being reported here.

MATERIALS AND METHODS

Plant Material

The matured fruits of Benincasa hispida were col-lected from Bangalore in the month of August and Sep-tember. Fruit was identified by the Botanist of Rural college of Pharmacy, Devanahalli. The voucher speci-men (BCSF) kept in our laboratory for future reference.

Extract Preparation

After removing skin and the seeds, the fruit pulp was dried under shade. The coarsely powdered fruit pulp was extracted successively with petroleum ether (B.P. 60-80°C), chloroform and methanol in a Soxhlet extrac-tor for 24-34 hrs. On evaporation of methanol from the methonolic extract in vacuo, a residue was obtained (yield 3.72% w/w) and was stored in desiccator. For pharmacological experiments weighed amount of the methanolic extract was suspended in 2% (w/v) aqueous tragacanth solution.

Animal Used

Albino Wistar of either sex weighing 160-180 g each were housed in standard metal cages. They were provided with food and water ad libitum. The rats were allowed a one-week acclimatization period before the experimental sessions.

Castor Oil Induced Diarrhea

The method followed here was the method of Awouters et al [ 13 ] with some modification. The origi-nal method has included only male Wister rats (220-250 g) and they were starved overnight before treatment with the selected drug in the next morning. In the pre-sent study (180-200 g) were fasted for 18 hrs. Animals were housed in five perforated steel cages containing six in each. None of the animal died even at an oral dose of 3.5 g/kg of BHFE. The doses of BHFE used were se-lected on a trial basis and was administered orally (200, 400, and 600 mg/kg) by gavage as suspension to three groups of animals. The fourth group received di-phenoxylate (5 mg/kg) orally as suspension as standard drug comparison. Fifth group, which served as control received 2% (w/v) aqueous tragacanth solution. One hour after treatment each animal received 1ml of castor oil orally by gavage and then observed for defecation. Up to 4^th hour after the castor oil challenge the presence of characteristic diarrheal dropping were noted in the transparent plastic dishes place beneath the individual rat cages.

Gastro Intestinal Motility Test [ 14 ]

Rats were fasted for 18hrs and place in 5 cages con-taining six in each. Each animal was administered orally with 1ml of charcoal meal (3% deactivated charcoal in 10% aqueous tragacanth). Immediately after that, the first three groups of animals were administered orally with the extract (BFHE) suspension (200, 400 and 600 mg/kg). The fourth group received atropine (0.1 mg/kg, i.p.), the standard drug for comparison. The fifth group was treated with aqueous tragacanth solution as control. Thirty minutes later, each animal was killed and the intestinal distance moved by the charcoal meal from the pylorus was cut and measured and expressed as a per-centage of the distance from the pylorus to the caecum.

PGE₂-Induced Enteropooling [14]

In this method, rats of the same stock as above were deprived of food and water for 18 hrs and were placed in 5 perforated cages with 6 animals per cage. The first three groups were treated with BHFE 200, 400 and 600 mg/kg, p.o). The fourth group was then treated with aqueous tragacanth suspension as mentioned earlier, which served as control. Immediately afterwards, PGE₂ was administered orally to each rat (100 µg/kg) in 5% v/v ethanol in normal saline. After 30 minutes each rat was killed and the whole length of the intestine from the pylorus to the caecum dissected out and the contents were collected in a test tube and the volume was meas-ured.

Statistical analysis was performed by Student’s ‘t’ test and in all the cases results are expressed as mean ± SEM.

RESULTS

Inhibition of Castor Oil-Induce Diarrhea

The extract (BHFE) like the standard antidiarrheal agent, dipheoxylate, inhibited significantly the fre-quency of defecation when compared to untreated rats ( Table 1 ). Both substances also reduced greatly the wet-ness of fecal droppings.

Effect on Gastro-Intestinal Motility

The extract decreased propulsion of the charcoal meal through the gastrointestinal tract when compared with the control group. Atropine reduced the motility of the intestine significantly ( Table 2 ).

Anti-Enter Pooling Activity

PGE₂ induced significant increase in the fluid vol-ume of rat intestine when compared with control ani-mals receiving only ethanol in normal saline and control vehicle. BHFE significantly inhibited PGE₂-induced enteropooling (Table 3).

DISCUSSION

Several studies have shown that prior administration with some plant extracts had a protective effect on the intestinal tract [ 15 - 17 ]. In the present study, the metha-nolic extract of fruit of Benincasa hispida (BHFE) that have not been studied so far, was evaluated for its antidiarrheal potential against castor oil induced diar-rhea, gastrointestinal motility in charcoal meal test and PGE₂ induced enter pooling in Albino Wistar rats. There has been a statistically significant reduction in the incident and severity of diarrhea produced in experi-mental animal models.

The methanolic extract of fruit of Benincasa hispida (BHFE) exhibited significant antidiarrheal activity against castor oil induced diarrhea in rats. The extract had a similar activity as diphenoxylate when tested at 200, 400 and 600 mg/kg and statistically significant reduction in the frequency of defecation and the wetness of the fecal droppings when compared to untreated con-trol rats (i.e., rats receiving neither BHFE nor di-phenoxylate but castor oil (only).

It is widely known that castor oil or its active com-ponent ricinoleic acid induces permeability changes in mucosal fluid and electrolyte transport that results in a hypersecretory response and diarrhea [ 18 , 19]. The ex-perimental studies in rats demonstrated a significant increase in the portal venous PGE₂ concentration fol-lowing oral administration of castor oil [ 20 ]. Ricinoleic acid markedly increased the PGE₂ content in the gut lumen and also caused on increase of the net secretion of the water and electrolytes into the small intestine [ 21 ]. The liberation of ricinoleic acid from castor oil results in irritation and inflammation of the intestinal mucosa, leading to release of prostaglandins, which stimulate motility and secretion [ 22 ]. Inhibitors of pros-taglandin biosynthesis delayed castor oil induced diar-rhea [ 13 ]. Based on the facts, it seems reasonable to suggest that the antidiarrheal effect of the BHFE may be due to the inhibition of prostaglandin biosynthesis.

The extract appears to act on all parts of the intes-tine. Thus, it reduced the intestinal propulsive move-ment in the charcoal meal treated model; at all doses of extract showed activity similar to that of atropine. Pre-vious study shows that activated charcoal avidly absorbs drugs and chemicals on the surface of the charcoal par-ticles thereby preventing absorption [ 23 ]. Thus, gastro-intestinal motility test with activated charcoal was car-ried out to find out the effect of BHFE on peristaltic movement. The results also show that the BHFE sup-pressed the propulsion of charcoal meal thereby in-creased the absorption water and electrolytes.

The extracts also significantly inhibited the PGE₂ induced intestinal fluid accumulation (enter-pooling). It has been shown that E type of prostaglandins cause di-arrhea in experimental animals as well as human beings [ 24 ]. Their mechanism has been associated with dual effects on gastrointestinal motility as well as on water and electrolyte transport [ 25 ]. PGE₂ also inhibit the absorption of glucose, a major stimulus to intestinal absorption of water and electrolytes [ 26 ]. These obser-vations tend to suggest that the BHFE at all tested doses reduced diarrhea by inhibiting PGE₂ induced intestinal accumulation of fluid.

The above observations suggest that BHFE in graded doses reduced diarrhea by inhibiting intestinal peristalsis, gastrointestinal motility and PGE₂-induced enteropooling. These inhibitory effects of BHFE sup-port the use of the Benincasa hispida in folk medicine; justify its use as non-specific antidiarrheal agent. Hence, BHFE, on preliminary studies can be claimed as a po-tential antidiarrheal agent, the underlying mechanism appears to be spasmolytic and anti-enteropooling prop-erty by which the fruit and/or its extract produced relief in diarrhea.

ACKNOWLEDGEMENT

Authors are sincerely thankful to our honorable founder secretary Mr. C. Basavaraja, Rural college of Pharmacy, Devanahlli, Bangalore, for providing all kinds of facilities.

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