Antidiarrhoeal Mechanisms of Kutajarishta

 

Prashant B. Shamkuwar *

Government College of Pharmacy, Thiba Palace, Ratnagiri (India).

*Corresponding Author E-mail: shamkuwarp@rediffmail.com

 

 

ABSTRACT:

Antidiarrhoeal effect of Kutajarishta, an ayurvedic formulation was evaluated with different agonist and antagonist in castor oil induced diarrhoea in mice. Antidiarrhoeal effect of Kutajarishta was decreased by Isosorbide dinitrite (nitric oxide donor) and Glibenclamide (potassium channel blocker). Yohimbine (α2 adrenergic receptor blocker) did not produce any significant effect on the antidiarrhoeal activity of Kutajarishta. The results obtained suggest that antidiarrhoeal effect of Kutajarishta was through potassium channels and nitric oxide pathway and not through α2 adrenergic receptors.

 

KEYWORDS: Kutajarishta, nitric oxide pathway, potassium channels, adrenergic receptors

 

 


INTRODUCTION:

Diarrhoea is a condition of passage of loose, watery stools with increased frequency.1 It involves decrease in the absorption of fluid, increased secretion and increase in the motility of the gastrointestinal tract. Potassium channels, nitric oxide pathway and α2 adrenergic receptors play an important role in the diarrhoea.2 Nitric oxide mediates multiple physiological functions in the gastrointestinal tract, including mucosal blood flow, maintenance of mucosal integrity, and maintenance of vascular tone. In epithelial cells of the gastrointestinal tract, a variety of K+ channels are expressed, allowing adaptation to different needs.3 Activation of the symphathetic innervations of the intestines results in inhibition of peristaltic activity and a reduction in tone.4 This inhibitory effect is mediated mainly by α2 adrenoceptors.5 Present study was aimed to investigate the involvement of potassium channels, nitric oxide pathway and α2 adrenergic receptors in antidiarrhoeal activity of Kutajarishta.

 

MATERIALS AND METHODS:

Drugs:

i) Kutajarishta Syrup – Shree Baidyanath Ayurved Bhawan (P) Ltd. ii) Isosorbide dinitrate – Sigma Chemicals Ltd. iii) Glibenclamide – Sigma Chemicals Ltd. iv) Yohimbine –  Sigma Chemicals Ltd.

 

Composition of Kutajarishta:

Each 10 ml of Kutajarishta contains i) Kutaja (3 gm), ii) Draksa (1.5 gm), iii) Madhuka Puspa (300 mg), iv) Gambhari (300 mg), v) Dhataki (600 mg) vi) Guda (3 gm), vii) Asav Base (q.s.).

 

Animals:

Swiss albino mice of either sex, weighing 20 – 25 gm obtained from VIPER, Pune, were used for the experiments. They were kept in standard environmental condition, fed standard food and water ad libitum. All experiments were performed after an overnight fast. The Institutional Animal Ethical Committee of Government College of Pharmacy, Aurangabad, Maharashtra, India (GCPA/IAEC/2011/235, 11/03/2011), approved the study.

 

Acute toxicity study:

Acute oral toxicity of Kutajarishta was studied as per revised OECD guidelines number 423. Kutajarishta was devoid of any toxicity up to 20 ml/kg in albino mice by oral route. Hence for further studies 2.5 ml/kg doses of Kutajarishta was used.

 

Antidiarrhoeal effect of Kutajarishta with Isosorbide dinitrate, Glibenclamide and Yohimbine on castor oil induced diarrhea:.

Groups of six mice each were treated as outlined below:

Group 1 (Control group): Distilled water 10 ml/kg, p.o.

Group 2 (Test group): Kutajarishta 2.5 ml/kg, p.o.

Group 3 (Test group): Isosorbide dinitrate 150 mg/kg, p.o. (given 30 min prior to the administration of Kutajarishta 2.5 ml/kg, p.o.)

Group 4 (Test group): Glibenclmide 1 mg/kg, p.o. (given 30 min prior to the administration of Kutajarishta 2.5 ml/kg, p.o.)

Group 5 (Test group): Yohimbine 1 mg/kg, s.c. given 30 min prior to the administration of Kutajarishta 2.5 ml/kg, p.o.)

 

Castor oil (0.2 ml/mouse) was administered to each mouse after 30 minutes. The animals were then placed under separate glass funnels, with the floor lined with blotting paper, for observation for 4 h. The parameters observed were: onset of diarrhoea, total weight of faecal output, total weight of wet faeces, total number of faecal output, and number of wet faeces.6, 7, 8        

 

Statistics:

The results of all experiments were reported as mean ± S.E.M. Statistical analysis was carried out using Student’s ‘t’-test. A level of significance of P < 0.05 was regarded as statistically significant.

 

RESULTS:

Effect of Isosorbide dinitrate, Glibenclamide and Yohimbine on antidiarrhoeal activity of Kutajarishta in mice.

All the mice in control group produced copious diarrhoea, after castor oil administration during the observation for 4 h. Kutajarishta decreased the onset of diarrhoea, number of wet stools, total no of stools, weight of wet stools, and total weight of stools.

 

Kutajarishta (2.5 ml/kg) produced 48.54% inhibition of diarrhoea. Kutajarishta (2.5 ml/kg) along with isosorbide dinitrate (150 mg/kg), glibenclamide (1 mg/kg) and yohimbine (1 mg/kg) showed 42.45%, 37.9% and 51.51% of inhibition of diarrhoea respectively.

.

Figure 1: Antidiarrhoeal effect of Kutajarishta with Isosorbide dinitrate, Glibenclamide and Yohimbine in mice

The ‘X’ axis represents the dose of the drug,

The ‘Y’ axis represents percent inhibition of diarrhoea.

KUT - Kutajarishta (2.5 ml/kg), ISDN - Isosorbide dinitrate (150 mg/kg), GLIB - Glibenclamide (1 mg/kg), YOHI - Yohimbine (1 mg/kg).

 

DISCUSSION:

Luminal potassium channels in the colonic surface cells act as an exit pathway for K+. It hyperpolarize the membrane voltage, thereby fueling electrogenic transport mechanisms such as Na+-coupled reabsorption of nutrients or luminal Cl secretion. They are involved in reabsorptive and secretory processes in colonic epithelial cells.9, 10 Antidiarrhoeal activity of Kutajarishta was decreased when given with potassium channel blocker drug, Glibenclamide. It indicates that antidiarrhoeal effect of Kutajarishta may be through potassium channels.

 

Nitric oxide (NO) is known to have a protective effect on the gastrointestinal tract. NO is responsible for helping to maintain the integrity of the gastric epithelium and the mucus barrier. NO is one of the mediators of the intestinal secretion and diarrhoea induced by castor oil. 11, 12 A NO donor, Isosorbide dinitrate, has reduced the antidiarrhoeal effect of Kutajarishta indicating the involvement of nitric oxide pathway in the antidiarrhoeal effect of Kutajarishta.

Sympathetic stimulation decreases blood flow to the gut as well as decreases secretions and general gut activity.13 The effect of sympathetic stimulation often only lasts a few minutes, and then an auto regulatory effect takes over restoring the blood supply to normal. However, this shutting off of the blood supply acts to increase flow to other areas of the body in time of stress.14, 15 Yohimbine, a specific α2 adrenergic receptor antagonist, did not produce any significant effect on the antidiarrhoeal activity of Kutajarishta. Thus α2 adrenergic receptors are not involved in the antidiarrhoeal effect of Kutajarishta.  

 

CONCLUSION:

Kutajarishta produced antidiarrhoeal effect may be through potassium channels, nitric oxide pathway, but not via α2 adrenergic receptors.

 

ACKNOWLEDGEMENTS:

The author expresses their gratitude to the Principal, Government College of Pharmacy, Aurangabad, for providing research facilities.

 

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Received on 03.04.2013       Modified on 15.04.2013

Accepted on 19.04.2013      © RJPT All right reserved

Research J. Pharm. and Tech. 6(5): May 2013; Page 550-552