Keywords

Castor Oil; Defecation; Diarrhea; Intestinal Movement

Introduction

Diarrhea is a state of abnormal bowel movement that can be explained by a situation in where an adult’s daily stool exceeds 300 g and contains 60–95% of water [1]. Diarrhea can cause severe dehydration that can lead to death. In fact, it is the second most common cause of death of baby and neonate in developing countries, and a death toll of about 2-4 million children is causing every year due to diarrhea, especially in African region [2].

Although Oral Rehydration Therapy (ORT) is found very helpful in the treatment of diarrheal diseases, the Oral Rehydration Solution (ORS) formula supplemented with cooked rice powder was more effective than commercially available ORS treatment [3]. Numerous plants with wide potentiality in the treatment of variety of disorders and diseases have been discovered but the enhancement of ORT efficiency after using of traditional medicinal plants has not been much applauded by scientists [4,5]. However, report from Martinez et al. [6] evident that herbal remedies are still relevant in the treatment of diarrhea by mothers in developing countries.

Albizia procera (A. procera) (Family: Fabaceae) is a tree widely distributed from Southeast Asia to Papua New Guinea and Northern Australia. Leaves are locally said as insecticidal [7] and bark is used for fish poison. Bark is also considered useful in pregnancy and stomachache and is given with salt as a medicine for water buffalo [8]. The ethanolic extracts of bark have found significant in the treatment of anti-HIV-1 diseased model [9]. The bark and leaf extracts of A. procera showed potent DPPH scavenging activity [10,11]. Other biomedical importance of A. procera especially the analgesic, antibacterial and CNS depressant activities of fraction of A. procera leaves [12] also taken in consideration before going for present study designation. Thus, this research has been conducted to provide a scientific basis for use of this plant in the diarrheal disease.

Methods

Collection and identification of plant material

Fresh leaves of the plants were collected in between March- April, 2012 and identified by Botany Department, University of Chittagong, Bangladesh. A voucher specimen with accession No. 3778 has been deposited in Bangladesh National Herbarium, Chittagong, Bangladesh.

Preparation of plant extracts

The leaves were left to dry under shade, grinded and extracted with methanol by cold maceration for 7 days at room temperature. The extract was then filtered off through a cotton plug and finally through filter paper. The filtrate was concentrated using vacuum rotary evaporator at 500°C. The percentage yields of extracts were calculated methanolic 20 g. All the extracts were stored in a refrigerator for further use. In the intra-cutaneous sutures group retrospective patient file research was not in the scope of the Dutch Human Bound Research and ethical approval was not required.

Animal

Albino mice (weighing 20-30 g) of both sex, were housed in standard metal cages. They were provided with food and water ad libitum, and allowed a one-week acclimatization period prior to the study. The equipment, handling and sacrificing of the animals were in accordance with the European Council legislation 87/609/EEC for the protection of experimental animals [13].

Oral toxicity tests

Fifteen Swiss albino mice (20-22 g) were randomly divided into nine groups of five animals each. The mice were fed on mice pellets and water ad libitum. The animals were starved for 12 h prior to testing. Eight doses of the extract (200–1000 mg/kg body weights) were administered by oral intubation to the animals. All animals were observed for 24 h and general symptoms of toxicity and mortality were recorded [14,15].

Castor oil-induced diarrhea

Castor oil induced diarrhea model was carried out using the method described by Shoba and Thomas [16]. The animals were screened initially by giving 1 ml of castor oil and those showing diarrhea were selected for the final experiment. Twenty five albino rats were randomly divided in to five equal group (n=5) divided in to control group, standard group and test groups. The control group received vehicle (1 ml/rat). The standard group received loperamide at the dose of 3 mg/kg orally [17]. The test group received aqueous extract of A. procera leaves 100 and 200 mg/ kg orally. Each animal were placed in individual cage, the floor of which was lined with bloating paper. The floor lining was changed for every hour. Diarrhea was induced by oral administration of 1.0 ml castor oil to each rat. 1 hour after the above treatment during an observation period of 4 h, the total numbers of feces excreted by the animals were recorded. A numeric score based on the stool consistency was assigned as follows: normal stool=1, semi-solid stool=2 and watery stool=3. The number of diarrheal feces and percentage of Inhibition of diarrheal feces were calculated [18-20]. Percentage inhibition was calculated as follows:

Magnesium sulphate induced diarrhea

A similar protocol as for castor oil induced diarrhea was followed. Diarrhea was induced by oral administration of magnesium sulphate (MgSO₄) at the dose of 2 g/kg to the animals 1 h after the vehicle (1 ml/mice) to the control group, Loperamide (3 mg/ kg) to the standard group and to the extract treated groups. All the administrations were carried out through oral route [21].

Statistics

The results were expressed as mean ± SEM and analyzed statistically to find out significance difference between control groups against each test group separately. Graph-Pad Prism 6 was used in this regard to complete the data analysis. We did One-way ANOVA along with Dunnet test comparison. The value of P<0.05 was considered statistically significant.

Results

Acute toxicity test

Oral administration of MEAP produced no visible signs of toxicity in the animals at the 250 and 500 mg/kg body weight of the rats. No mortalities were recorded. In addition, no toxic symptoms were observed and neither food nor water intake was found to be reduced during the period.

Castor oil induced diarrhea

The doses of MEAP significantly decreased (P<0.05) the total number of diarrheal feces produced by administration of castor oil (3.00 ± 0.58 at the dose of 200 mg/kg) as compared to castor oil treated control group (7.67 ± 1.45) and comparable to the standard drug. The percentage of inhibition of castor oil induced diarrhea in MEAP treated rats was 60.89% at the dose of 200 mg/ kg body weight of the rats and presented in Table 1.

Table 1: Effect of methanolic leaves extract of Albizia procera on castor oil induced diarrhea in mice.

MgSO₄ induced diarrhea

The doses of MEAP significantly decreased (P<0.05) the total number of diarrheal feces produced by administration of MgSO₄ (2.33 ± 0.33 at the dose of 200 mg/kg) as compared to MgSO₄ treated control group (6.67 ± 1.45) and comparable to the standard drug. The percentage of inhibition of castor oil induced diarrhea in MEAP treated mice was 65.07% at the dose of 200 mg/kg body weight of the mice and presented in Table 2.

Table 2: Effect of MgSO₄ induced leaves extract of Albizia procera on castor oil induced diarrhea in mice.

Discussion

Diarrhea is usually considered a result of altered motility and fluid accumulation within the intestinal tract. Many antidiarrheal agents act by reducing the gastrointestinal motility and/or the secretions. Castor oil causes diarrhea due to its active metabolite, recinolic acid [3] which increases peristaltic activity in the small intestine leading to changes in the electrolyte permeability of the intestinal mucosal membrane. The precise mechanism of action of castor oil is through elevated prostaglandin biosynthesis [22,23]. Prostaglandin contributes to the pathophysiological functions in gastrointestinal tract [24]. Inhibitors of prostaglandin biosynthesis delay castor oil-induced diarrhea [25].

Castor oil causes diarrhea due to its active metabolite, ricinoleic acid, which is liberated as a result of action of lipases on castor oil. This stimulates peristaltic activity in the small intestine, leading to changes in the electrolyte permeability of the intestinal mucosa. It also stimulates the endogenous prostaglandins [23]. Castor oil elicits secretory and motility diarrhea [26]. Inhibitors of prostaglandin synthesis are known to delay diarrhea induced with castor oil [18]. The observations suggest that the antidiarrheal effect of the extract may be due to inhibition of prostaglandin synthesis. The extract also exhibited a significant inhibition of the small intestine propulsive movement the effect was comparable to that of the standard drug Loperamide, used in the study. Anti-diarrheal and anti-dysenteric of medicinal plants were found to be due to the presence of tannins, alkaloids, saponins, flavonoids, steroids and terpenoids [27]. Further studies are required to confirm the underlying mechanism of the observed activity of the plant.

Wet stool is one of the major characteristics of diarrhea, which is usually caused by altered motility and fluid accumulation in the intestine lumen. MgSO₄, an osmotic-acting laxative, has been reported to induce diarrhea by increasing the volume of intestinal content through prevention of reabsorption of water. In the present study, diarrhea characterized by intestinal fluid accumulation was developed in mice after the administration of MgSO₄. Obvious watery stool in the colon, increased fecal water content, and increased defecation were observed in vehicle-treated mice. While in MEAP-treated mice we noticed that MEAP attenuated the severity of diarrhea by reducing the degree of watery stool in the colon, the fecal water content, and in a dose-dependent manner. The results suggest that MEAP has an antidiarrheal activity in MgSO₄-induced diarrhea in mice partly via reducing the colonic water secretion induced by MgSO₄.

The antidiarrheal activity of the plant extract was not comparable to the standard drug, loperamide, which at present is one of the most efficacious and widely employed antidiarrheal drugs. Loperamide effectively antagonizes diarrheal activity induced by castor oil [28], prostaglandins [29] or cholera toxin [30]. Loperamide, apart from regulating the gastrointestinal tract, is also reported to slow down transit in the intestine, reduce colon flow rates and consequently any effect on colonic motility [31,32]. The antimuscarinic drug, atropine and different doses of the extract decreased the propulsive movement in the charcoal meal study. This is possible due to its anticholinergic effect [33]. The significant inhibition of the castor oil and MgSO₄-induced enteropooling in mice suggests that A. procera leaf extract produces relief in diarrhea by spasmolytic activity in vivo and also anti-enteropooling effects [34].

Conclusion

In conclusion, the present study revealed that Albizia procera contains pharmacologically active substances effective for management of diarrhea. Further studies are required to fully investigate the mechanisms responsible for this observed antidiarrheal activity.

Acknowledgement

All authors are very much appreciative to lab staffs of Department of Pharmacy, International Islamic University Chittagong for their unforgotten able help and assistance and also thankful to Rafikul Islam, Assistant Professor, and IIUC.

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