Evaluation of Antidiarrhoeal Activity Cleome gynandra Leaf Extracts on Magnesium Sulphate- and Castor Oil-Induced Diarrhoea in Wistar Rats

Nadiminti Satish Reddy*, D. Santhosha, Vidyasabbani, P. Vishwanath Reddy, Ch. Venu

Department of Pharmacology, Vishnu Institute of Pharmaceutical Education and Research, Vishnupur, Narsapur, Medak, Telangana, India.

*Corresponding Author E-mail: satishreddy949@gmail.com

ABSTRACT:

The methanolic extract leaves of Cleome gynandra (Family Capparidaceae) was selected to evaluate the antidiarrhoeal activity against castor oil and magnesium sulphate induced diarrhoea in wistar albino rats. The present study was performed at three different doses of 100 mg/kg ,200mg/kg and 400mg/kg methanolic extract of plant .The Methanolic plant extracts shows more effective antidiarrhoeal activity against castor-oil induced diarrhea compared to magnesium sulphate induced diarrhoea in a dose dependent manner .From the results we conclude that methanolic leaf extracts of Cleome gynandra had reported antidiarrhoeal activity similar to standard drug loperamaide at dose of 3 mg/kg .

KEYWORDS: Cleome gynandra, antidiarrhoeal activity, Loperamide, Castor oil, Magnesium sulphate.

INTRODUCTION:

Diarrheal diseases are a major problem in Third World countries and are responsible for the death of millions of people every year¹. Diarrheoa is an alteration in normal bowel movement and is characterized by an increase in the water content, volume, or frequency of stools². Plants have long been a very important source of new drugs. Many plant species have been screened for substances with therapeutic activity. Medicinal plants are a promising source of ant diarrheal drugs³. For this reason, international organizations including the World Health Organization (WHO) have encouraged studies pertaining to the treatment and prevention of diarrheal diseases using traditional medical practices^4,5,6. Cleome gynandra Linn. (Capperdiceae) is described in Ayurveda and other system of medicine as a curative medicine for neuralgia, headache, cough, wounds, anthelmintic, rubefacient, counterirritant and for snake bite and scorpion sting etc⁷.

Various species of cleome are used medicinally in Indo-China, Philippines, Island, North and Central America. The leaf paste of plant has been used in rheumatism, neuralgia, headache and stiff neck. Its warm juice is a popular remedy for ear disease. The leaf juice is applied in skin disease. Juice of fresh leaves is applied externally during pyorrhea and it is also used as a wormicide⁸. Hexacosanol, â-D-glucoside of â sitosterol, free â sitosterol and kaempferol have been isolated from the seed of Cleome gynandra⁹. It also contains the minor components 5, 7- dihydroxychromone, 5-hydroxy-3, 7, 4-trimethoxyflavone and luteolin¹⁰. The present study was undertaken to find out the antidiarrhoeal activity methanolic leaf extracts of Cleome gynandra against castor oil and magnesium sulphate induced diarrhoea in wistar albino rats .

MATERIALS AND METHODS:

Plant Material and Preparation of Extract:

The leaves of Cleome gynandra were collected in the month of August from Department of Botany, Tirupati district authenticated by Prof. Madhav Shetty (Voucher no:492) . The plants were air dried in shade for 15 days and then the aerial parts of the plants taken for the study. Coarse powder of Cleome gynandra was extracted separately with 70% v/v , Methanol using a soxhlet apparatus. The extracts thus obtained were dried under reduced pressure at a room temperature not exceeding 40⁰c to get the extracts. The percentage of yield was found to 10%.

Animals:

Albino Wistar rats (weighing 150-200 g) of both sexes, were procured from NIN , Hyderabad, India and were housed in standard metal cages. They were provided with food and water ad libitium, and allowed a one week acclimatization period prior to the study. The protocol was approved by Institutional animal ethical committee of VIPER, Narsapur Medak and the study was performed according to the CPCSEA guidelines 1358/ERe/S/10/CPCSEA.

Preliminary acute toxicity test:

The methanolic leaf extract of Cleome gynandra was administered orally in doses of 125, 250, 500,1000, 2000mg/kg body weight to animal groups (one dose per group). Simultaneously, the control animals received normal saline (5ml/kg). The general signs and symptoms of toxicity, intake of food and water and mortality were recorded for a period of 48 h and then for a period of 14days.

Experimental Procedure for Antidiarrhoeal Activity:

Healthy Wistar rats were distributed into 5 groups, each group consisting of 5 animals, which received the treatments in following manner

Group I:

Normal control [1% carboxymethyl cellulose (CMC) 10 mL/kg ,p.o

Group II:

Standard drug : loperamide 3 mg/kg ,P.o

Group III:

Methanolic leaf extract of Cleome gynandra (100 mg/kg),P.o

Group IV:

Methanolic lraf extract of Cleome gynandra (200 mg/kg),P.o

Group v:

Methanolic leaf extract of Cleome gynandra (400 mg/kg), P.o All animals were initially screened for induction of diarrhoea by administering 2 mL of castor oil or 2 g/kg body weight dose of magnesium sulphate. Only animals which developed diarrhoea were selected for antidiarrhoeal studies.

Castor Oil-induced and Magnesium Sulphate-induced Diarrhoea in Rats^11-13.

Albino Wister rats of both sex were taken and weiged, 160 to 200 g were taken and kept fasting for 24 hours with access to water . The standard anti-diarrhoeal drug (loperamide, Yashica Pharmaceuticals Ltd., Thane, Maharashtra, India) and test sample (Methanolic extract of Cleome gynandra, 100, 200, 400 mg/kg body weight) to be tested were administered orally by gavage. For castor oil-induced diarrhoea, 2mL of castor oil was administered orally to each animal by gavage after one hour after administration of drug/extract. For magnesium sulphate-induced diarrhoea, magnesium sulphate was administered at a dose of 2 g/kg orally to each animal, 40 minutes after administration of drug/extract. All animals were placed in individual cages, where floor was lined with non-wetting paper. Non-wetting paper were changed every hour up to 5 hours. Characteristic diarrhoeal droppings of every hour up to the 5th hour were recorded after draining the urine by gravity, the stools were taken and calculated. A numerical score based on stool consistency was assigned. Normals tool was assigned as 1, semi-solid stool as 2 and watery stool as 3.Mean of diarrhoeal droppings passed by treatment groups was compared to control group

Statistical analysis:

Data were analyzed by one-way ANOVA followed by Dunnett’s t-test. P-value<0.01was considered significant and results were expressed as mean ± SD.

RESULTS AND DISCUSSION:

In castor oil-induced diarrhoea model, Methanolic extract of Cleome gynandra showed antidiarrhoeal effect in Wistar rats. Loperamide, the standard antidiarrhoeal drug, was same in reducing the number of faeces by 70.94%, while MeOH extract was found to be most effective, reducing diarrhoeal droppings by 70.90% at 400mg/kg . MeOH extract significantly (p<0.01) at a dose of. reduced the wet faeces and total number of faeces, when compared to control group and we conclude that Cleome gynandra had reported antidiarrhoeal activity in dose dependent manner. (Table1).

Table 1: Effect of MeOH Cleome gynandra on castor oil-induced diarrhoea in Wistar rats.

Treatment	Dose (mg/kg, p.o.)	Mean of wet faeces in 6 hours (n)	Mean of total number of faeces in 6 hours (n)	Feaces reduction (%)
Control	10 ml	20.62±1.02	23.4±2.07	---
MeOH extract	400	5.6 ±0.80	6.8±0.80* *	70.90**
MeOH extract	200	9.84 ±0.65	13.2±1.92*	46.77*
MeOH extract	100	19.80 ± 0.73	12.20 ± 1.06*	44.40*
Loperamide	3	5.62±0.90	6.8±0.84* *	70.94 **

** indicates p<0.01 highly significant

* indicates p<0.005 less significant

Table 2: Effect of MeOH Cleome gynandra on magnesium sulphate diarrhoea in Wistar rats.

Treatment	Dose (mg/kg, p.o.)	Mean of wet faeces in 6 hours (n)	Mean of total number of faeces in 6 hours (n)	Feaces reduction (%)
Control	10ml	20.62±1.02	23.4±2.07	---
MeOH extract	400	4.62 ±0.47	9.4±1.34	62.1**
MeOH extract	200	10.38±0.88	14.0±1.87	40.17*
MeOH extract	100	19.80 ± 0.73	12.20 ± 1.06	34.40*
Loperamide	3	5.62±0.90	6.8±0.84	71.77**

** indicates p<0.01 highly significant

* indicates p<0.005 less significant

CONCLUSION:

The results of this investigation revealed that methanolic leaf extract of Cleome gynandra showed significant ant diarrheal properties in dose dependent manner in both the models. These attributes may provide the rationale for the use of Cleome gynandra in diarrheoa management by traditional healers. Further research is needed to fractionate the MeOH extract and isolate the molecule(s) responsible for the antidiarrheal activity observed.

ACKNOWLEDGEMENT:

We would like to thank our Chairman and Dr. A. Ramesh, Principal, VIPER, Narsapur, Medak for providing the facilities. And also Professor. Madhav Shetty, Department of Botany, Tirupati for the botanical identification and collection of the plant.

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Received on 05.10.2015 Modified on 16.10.2015

Res. J. Pharmacology & P’dynamics. 7(4): Oct.-Dec., 2015; Page 184-186

DOI: 10.5958/2321-5836.2015.00037.3