Antidiarrhoeal Activity of Bark Extracts of Caesalpinia sappan. Linn.

Rasheed Ahmed, KL Senthilkumar* and M Rajkumar

Department of Pharmacognosy, Padmavathi College of Pharmacy, Periyanahalli -635205, Dharmapuri, Tamilnadu.

ABSTRACT

The antidiarrhoeal activity and gastrointestinal motility reducing activity of alcoholic and aqueous extracts of bark of Caesalpinia sappan,linn. were evaluated at two dose levels. Both the extracts showed significant antidiarrhoeal activity and reduced the mean weight of faeces and reduced the gastrointestinal motility significantly.

INTRODUCTION

Caesalpinia sappan, Linn. (Fabaceae) locally known as sappan-wood found almost throughout India mainly in TamilNadu and Kereala.¹ The bark used for tuberculosis, diarrhea, dysentery, postpartum tonic.² However, there is no scientific proof justifying the traditional use of bark in the treatment of diarrhoea. Hence, the present work was undertaken to evaluate its potential antidiarrhoeal efficacy in Swiss albino Mice.

MATERIAL AND METHODS

Plant Material

The bark of Caesalpinia sappan, Linn. were collected in the month of August 2008 from the ABS Botanical Garden, Salem-Dist, TamilNadu. The species for the proposed study was identified and authenticated as Caesalpinia sappan (Linn.). by Dr. P. Jayaraman, Botanist, Plant Anatomy Research Center (PARC), Chennai.

Preparation of Extracts

Preparation of the different extracts of Caesalpinia sappan, linn. powdered bark is done successively in a continuous soxhlet extractor with the following solvents. Hexane, Chloroform, Ethyl acetate, Alcohol and distilled Water. The yield of alcoholic and aqueous extracts was found to be 26.89% and 17.58% w/w respectively. Both the extracts revealed the presence of alkaloids and tannins. The extracts were stored in desiccators and used for further experiments.

Animals Used

Swiss Albino mice for antidiarrhoeal activity and Wister rats of female sex weighing 140-180gms were used for acute toxicity. Animals were maintained under standard laboratory conditions. Study protocol was approved from the Institutional Animals Ethics Committee (IAEC).

ANTIDIARRHOEAL ACTIVITY

Castor oil Induced Diarrhoea

In the present study animals were divided into six groups of six mice in each group. The animals were divided into control, positive control and test groups containing six mice in each group. Control group received vehicle (1% Tween 80 in water) at a dose of 10 ml/kg body weight orally. The positive control group received loperamide at the dose of 3 mg/kg orally, and test groups received the Alcoholic extract and aqueous extract at the doses of 200 and 400 mg/kg body weight orally. Each animal was placed in an individual cage, the floor of which was lined with blotting paper. The floor lining was changed every hour.

Treatment	Dose (mg/kg)	Total number of faeces in 4 hrs	Total number of weight faeces in 4 hrs	%Inhibition
Control	-	13.67±0.99	8.67±0.49	-
Loperamide	3	3.17±0.48*	1.83±0.31*	78.89
Alcohol Extract	200	9.66±0.51*	5.67±0.67	34.60

	400	4.25±0.34*	2.83±0.60*	67.35
Aqueous Extract	200	11.5±0.56	7±0.58	19.26
Aqueous Extract	400	7.83±0.60*	4.5±0.43*	48.09

TABLE NO. 1: Effect of bark extracts on castor oil-induced diarrhoea in mice

Results are mean±S.E.M. n=6. * Significantly different from the control at P<0.001

TABLE NO. 2: Effect of bark extract on the intestinal transit of charcoal meal in mice

Treatment	Dose (mg/kg)	% Movement of charcoal meal	% Inhibition
Control	-	65.5±2.28	-
Atropine sulphate	0.1	27.17±2.10*	58.51
Alcohol Extract	200	49.67±1.45*	24.16
Alcohol Extract	400	28.5±3.06*	56.44
Aqueous Extract	200	56.83±2.06	13.23
Aqueous Extract	400	42.5±1.82*	35.11

Results are mean±S.E.M. n=6. * Significantly different from the control at P<0.001

Diarrhoea was induced by oral administration of 0.5 ml castor oil to each mouse, 30 min after the above treatments. During an observation period of 4 hours, the total number of faecal output and the number of diarrhoeic faeces excreted by the animals were recorded.^3-5

Gastrointestinal Motility Test:

For gastrointestinal motility test, animals were divided into six groups of six mice in each group. The animals were divided into control and test groups containing six mice in each group. Control group received vehicle (1% Tween 80 in water) at a dose of 10 ml/kg body weight orally. Positive control group received atropine sulphate at the dose of 0.1 mg/kg intraperitoneally, and test groups received the alcohol extract and aqueous extract at the doses of 200 and 400 mg/kg body weight orally respectively. After 30 min, mice of each group were fed with 1ml of charcoal meal (3% suspension of deactivated charcoal in 0.5% aqueous methyl cellulose). After 30 min of the administration of charcoal meal, the animals of each group were sacrificed and the length of the intestine (pyloric sphincter to caecum) as well as the distance travelled by charcoal as a fraction of that length was measured. The charcoal movement in the intestine was expressed as a percentage.^6-8

Statistical Analysis:

The results are expressed as mean ± SEM of six independent experiments. Statistical significance between group was evaluated by one-way analysis of variance (ANOVA) followed by Dunnett’s test. A P < 0.001 value was considered as statistically significant.

RESULTS AND DISCUSSION:

In the castor oil-induced diarrhoeal experiment in mice, the alcoholic and aqueous extracts at the doses of 200 and 400 mg/kg, reduced the total number of faeces as well as the total weight of diarrhoeic faeces in a dose dependent manner (Table no.1). These results were shown to be statistically significant. In the gastrointestinal motility test, the both extract, at the doses of 200 and 400 mg/kg, retarded the intestinal transit of charcoal meal in mice when compared to the control (Table no.2). Several mechanisms have been previously proposed to induce the diarrhoeal effect of castor oil. These include inhibition of intestinal Na⁺,K⁺-ATPase activity to reduce normal fluid absorption, activation of adenylate cyclase or mucosal cAMP mediated active secretion stimulation of prostaglandin formation, platelet activating factor and most recently nitric oxide has been claimed to contribute to the diarrhoeal effect of castor oil. Despite the fact that these numerous mechanisms have been proposed, it has not been possible to define castor oil’s correct mechanism of action. However, it is well documented that castor oil produces diarrhoea due to its most active component recinoleic acid by a hypersecretory response. Since the alcoholic and aqueous extract at dose 400 mg successfully inhibited the castor oil-induced diarrhoea, the extract might have exerted its antidiarrhoeal action by antisecretory mechanism. This was also evident from the reduction of total number of wet faeces in the test groups in the experiment.

The extract may have increased the absorption of water and electrolyte from the gastrointestinal tract, since it delayed the gastrointestinal transit in mice as compared to the control. The delay in the gastrointestinal transit prompted by the extracts might have contributed, at least to some extent, to their antidiarrhoeal activity by allowing a greater time for absorption. The alcoholic extract showed more significant activity as compared to aqueous extract.

REFERENCE:

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2. Article taken from www.botany.hawaii.edu

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Received on 10.04.2009

Accepted on 04.06.2009

Research J. Pharmacology and Pharmacodynamics 1(3) Nov - Dec. 2009; 1(3): 128-129