International Journal of Phytomedicine 3 (2011) 68-74

Original Research Article ISSN: 0975-0185

Antidiarrhoeal activity of leaf extract of Moringa Oleifera in experimentally
induced diarrhoea in rats
Lakshminarayana M1*, Shivkumar H2, Rimaben P2, Bhargava VK2.

*Corresponding author: Abstract
To evalauate the antidiarrhoeal activity of the hydroalcoholic extract of
Lakshminarayana M moringa oleifera leaves. The hydroalchoholic extract was evaluated using
1 Department of rodent animal models of diarrhoea like the castor oil and magnesium sulfate
Pharmacology, induced gastrointestinal motility, in a model of enteropooling induced by the
SCS College of Pharmacy, administration of castor oil and PGE2, Charcoal meal test. Acute toxicity
Harpanahalli-583131, and phytochemical constituents were also been evaluated using standardized
Karnataka, India. methods. The results of the present study indicates that the hydroalcoholic* extract of moringa oleifera leaves was effective in inducing a significant
2 Krupanidhi College of protection against experimentally induced diarrhoea by castor oil and
pharmacy, Bangalore magnesium sulfate, as evidenced by a decrease in the number of frequency,
Karnataka, India. weight of stools after 4 hours with respect to control. The extract also prevented the enteropooling induced by castor oil and PGE2 at all the doses
tested. Acute toxicity studies indicated that the extract is safe till 2500
mg/kg. The antidiarrhoeal activity though, not ascribed to any particular
phytochemical present, general tests performed indicated the presence of
flavonoids, tannis which were reported to produce antidiarrhoeal activity.
These results showed that Moringa oleifera leaves possess anti-diarrheal
properties mediated through inhibition of hyper secretion and
gastrointestinal motility that substantiate its use in the treatment of diarrhea
in traditional medicines or folklore use.
Keywords: enteropooling, hyper secretion, Moringa oleifera.

Introduction These bacterial are commonly infested either by
Diarrhoea is characterized as rapid movement of means of polluted water or consumption of
faecal matter through intestine resulting in poor contaminated food and by physical contact like
absorption of water, nutritive elements and handshake etc.
electrolytes producing abnormal frequent
evacuation of watery stools. According to world These bacteria cause the influx of water and ions
health organization, it is the one of the most to the intestinal lumen and thus increase the
common cause of morbidity and mortality in intestinal motility, thereby causing watery stools.
many developing countries effecting mainly the Such secretory diarrhoea is treated by the
infants and children’s [1]. It is often caused by administration of oral rehydration salts in
enterotoxins which are produced by bacteria such children or adults to reduce the loss of essential
as Escherichia coli, Salmonella typhi, Salmonella electrolytes and maintain the body fluids
typhimurium, Clostridium difficile, Clostridium osmolality [2]. Alternatively, many opioid drugs
freundii, Aeromonas hydrophila, Campylobacter like Diphenoxylate, Loperamide, Diloxanide
jejuni and Vibrio cholerae, to name a few [2]. furoate for protozoal infections induced diarrhea
This work is licensed under a Creative Commons Attribution 3.0 License.

anti-diabetic [10]. present days there has been great interest in extracted with circulating 70% hydroalcohol at herbal remedies for the treatment of such 800C. extract prepared thus was administered to mice antioxidant [6]. 250 and 500 mg/kg were of Moringa oleifera as an antidiarrhoeal. racecadotril. the present study was defecation etc. Karnataka. vegetable. Ethanol (Merck. In view of this. 69 .25 gm (22. the study. asthmatic [11]. Lakshminarayana et. But all of the existing drugs suffer from adverse effects like the Preparation of crude extracts and induction of bronchospasm. Mumbai). external sores and ulcers.S. flavonoids. For this reason. leaves. K. for concentrating. undertaken to justify the traditional use of leaves Accordingly 100. coarsely and loaded onto a soxhlet evaporator. The second group of Lopermaide. purposes such as medicine. the obtained ailments. tree has been used in folk medicine for the Tests for glycosides and steroids revealed their treatment of various diseases such as urinary tract absence in the obtained hydroalcoholic extract. anti-ulcer [7]. All the parts of the alkaloids. method in accordance to OECD guidelines [15]. Later. the standard drug (Micro labs.R. department of Pharmacognosy. hepatoprotective [12] and anti. The extract was found to posse’s cooking and as a cosmetic. Prabhu. Chemicals: Ketamine (Themis medicare. Animals: Albino mice (25-30 g) and Wistar rats Materials and methods: (200-250 g) were used in the present study. are available in (Astra zeneca. All Plant material: the animals were housed in polypropylene cages Fresh leaves of Moringa oleifera were collected and maintained in a standardized animal house in and around Davangere. diabetes. as spice for Khandalwal14. literature between 25 to 35 g were used for the study. intestinal obstruction and The shade dried plant material was powdered constipation by loperamide [3]. anti-cancer [8]. After review failed to offer any scientific validation on oral administration of extracts various parameters the antidiarrhoeal activity of Moringa oleifera like body temperature. 15. College of pharmacy. anti. acacia. lopermaide. Concentration of the extract produced many remain to be evaluated scientifically. All the tests were deposited at the Museum of SCS College of performed according to OECD guidelines [15] pharmacy.6% W/W) yield. relative humidity. castor oil. the market for treating diarrhoea. Preliminary phytochemical screening of the obtained extract Moringa oleifera (family: Moringaceae) is one was performed in accordance to the tests among such plants cultivated for different described elsewhere by Kokate and K. cancer. Charcoal meal. animals received the standard drug. International Journal of Phytomedicine 3 (2011) 68-74 and dysentry. Magnesium First group always served as vehicle or control sulfate (SD Fine chemicals. and institutional animal ethics committee approved the study protocol. HIV-aids. Although several medicinal plants have extract was flash evaporated at 500C and used for gained importance for the treatment of diarrhoea. However. group in all the cases. Voucher under standard conditions of temperature (24 ± herbarium specimens have been authenticated by 2ºC). Uttarakhand). Healthy adult female Swiss albino mice weighing hypertensive properties [13]. Harapanahalli and and water ad libitum. and 12 hr light/dark Prof. Animals were fed with standardized chow S.C. and tannins. micturation. for LD50 determinations using the fixed dose anti-hyperlipidaemic [9]. al. infections. gastritis etc [4]. Mumbai) and PGE2 blockers like atropine sulfate etc. selected for treatment during the study. CNS activity. The plant has Dose selection and treatment: Hydroalcoholic been reported to possess anti-inflammatory [5]. carbohydrates. muscarinic receptor bangalore). Bangalore). Gum Five groups of animals were used for the study. were observed for 24 h. vomiting by characterization: racecadotril. cycle.

250 Immediately after extract administration. al. The extract was suspension. The second group received standard administered orally at 100.M of n =6 After 30 min. Lakshminarayana et. 1 mL of Castor oil was administered induced diarrhoea model. at the doses mentioned above. The oral route. The extract was administered orally at 100. divided into six groups of six in each group. 60 min after drug administration. following administration of castor animals. One group served as negative Rats of either sex were used in the study and control and the other one as a positive control. was administered with PGE 2 was administered with aqueous 2% acacia (100 μg/kg p. the animals in each group received drug treatment was done. six groups of enteropooling [19]: animals were used. each of these animals were given faecal material and the frequency of defecation 1mL of charcoal meal (3% charcoal suspension in were noted up to 4 hr in the transparent metabolic 5% suspension of acacia) by oral route. gum as the suspending agent. 70 . all rats were sacrificed by overdose of of variance and Dunnet`s posttest. For prostaglandin (PGE2) Prostaglandin E2 (PGE 2) induced induced enteropooling model. the stomach the base. All cages with pre weighed plastic dishes placed at animals were sacrificed after 30 min. loperamide (3 mg/kg) orally as suspension.m. After 30 min following group respectively.05 were considered significant.) only. One hour later after magnesium sulphate (2 g/kg) orally. previously. The second group. The first group. 250 and 500 mg/kg to drug. which served as negative control was Castor oil induced diarrhoea [16]: administered with vehicle (2% acacia suspension) Rats of either sex (150-250gm) were fasted for 18 and1 ml of 5%v/v ethanol and normal saline by h. Values with p ketamine. fourth and fifth was administered. third. on a clean glass surface. fourth and fifth group respectively. Mean values were evaluated by Analysis oil. They were divided into four groups (n=6). Values are expressed as mean ± S. pre weighed plastic dishes placed at the base. its content collected in up to 4 h in the transparent metabolic cages with measuring cylinder and volume measured. Again. After 60 min of drug administration of PGE 2 each rat was sacrificed treatment. 1 mL of castor oil was administered to all animals Statistical analysis: including the control or vehicle treated group. Albino Wistar rats were treated as described earlier except that atropine sulfate was used as a Magnesium sulphate induced diarrhoea [17]: standard drug and administered at a dose of 5 Animals and treatment was similar to castor oil mg/kg i. whole length of the intestine from < 0. which served as control positive control.o. the animals of each group received by administering excessive dose of ketamine and 1ml of castor oil orally and the watery faecal the whole length of the intestine from pylorus to material and frequency of defecation was noted caecum was dissected out. Weight of plastic dish before and after defecation Effect on Charcoal meal test [20]: was noted and compared to control. its content Table 1: Effect of hydroalcoholic extract of Moringa oleifera (HMO) on Castor Oil Induced Diarrhoea in Rats. International Journal of Phytomedicine 3 (2011) 68-74 All the other three groups of animals received the collected in measuring cylinder and volume hydroalcoholic extract in suspension using acacia measured. the drug treatments. The distance moved by the charcoal meal from the pylorus was measured Castor oil induced enteropooling [18]: and then expressed as a percentage of the Treatment remained the same as mentioned distance from the pylorus to the caecum. pylorus to caecum was dissected out. Weight of plastic dish before and after and small intestine were removed and extended defecation was noted and compared to control. PGE2 and 500 mg/kg dose to third. After 60 min of drug orally to rats and left for one hour after which treatment. which served as first group of animals.E.

The values are mean ± SEM.00 HMO.6±0.083*** 50.3±0.28±0.2* 6.044*** 74.6±0.54*** 0. among all the extracts being extract administered as shown in Tables 3 and 4. International Journal of Phytomedicine 3 (2011) 68-74 Groups Treatment Dose Mean frequency of Mean wt of fecal Mean wt of faeces % Mg/kg diarrhoea ± SEM drops ± SEM ± SEM after 4hr protection I Control 1 mL 3.00 II Loperamide 3 0.. n=6. *p<0.2±0.778 1.4±0. the reason for which could not be explained.6±0.48 11.26±0. Groups Treatment Dose Mean frequency of Mean wt of fecal Mean wt of faeces % mg/kg diarrhoea ± SEM drops (g) ± SEM (g) ± SEM after 4hr protection I Control 1 mL 4. n=6.27 IV HMO 250 1.050*** 87.050*** 46. The faecal drops and means weight of faeces with low dose of the extract produced more inhibition respective to control as shown in Table 1 and 2.4** 6±0.54±0. 71 . n=6.6±0.92*** 0.81*** 81.44*** 0.42 V HMO 500 1. Hydroalcoholic extract of Moringa oleifera.gram. mean number of anti enteropooling effect of Moringa oleifera. **p<0.35 mL.0±0. Castor oil and PGE2 failed to produce In a model of castor oil and magnesium sulfate enteropooling in animals at all doses of the induced diarrhoea.01 and ***p<0.gram.001 vs control. ***p<0. Table 2: Effect of hydroalcoholic extract of Moringa oleifera (HMO) on Magnesium sulfate induced Diarrhoea in Rats.67*** 0.4±0.82±0.058 0.05*** 53. of enteropooling than the other doses tested in PGE2 model of diarrhoea.14±0.94 III HMO 100 1.5*** 2.4±1.6±0.37*** 0.0±0.01 and ***p<0. **p<0.7±0.001 vs control. Table 3: Effect of hydroalcoholic extract of Moringa oleifera (HMO) on castor oil induced Enteropooling in Rats.00 IV HMO 250 1. *p<0.05*** 90.. the higher dose produced a significant The protective effect of the extract indicates.033*** 86.2*** 1.72 III HMO 100 2.154±0.4±0.6±0. the decrease in mean frequency.044*** 0.001 vs control.2±0.24*** 3.05.13 HMO.12 0.44 V HMO 500 1.34±0.42 III HMO 100 1.80*** 80.17 V HMO 500 1.2** 5±0.07*** 48.05.0±0. Groups Treatment Dose mg/kg Mean volume of intestinal fluid (ml) ± SEM % protection I Control 1 mL 2.12±0. Lakshminarayana et.00 II Loperamide 3 0.24*** 1.021 10.2±0.8±0. The values are mean ± SEM. effective.070*** 75.16±0.6±0.24 1.6±0.83*** 0..2±0. al.54±0. The values are mean ± SEM. g.00 II Loperamide 3 0.6±0.31*** 2.031*** 0. g.5*** 0. Results: Both. Millilitres. Hydroalcoholic extract of Moringa oleifera.05*** 91.09 IV HMO 250 0.

85±0. International Journal of Phytomedicine 3 (2011) 68-74 Table 4: Effect of hydroalcoholic extract of Moringa oleifera (HMO) on PGE2 induced Enteropooling in Rats.26±2. counteracted the increase in electrolyte secretion by means of an anti-electrolyte permeability Castor oil or its triglyceride is hydrolyzed by action. The antidiarrhoeal Moringa oleifera mechanism of action in activity of the extract against experimentally inhibiting diarrhea involves anti propulsive and induced diarrhoea by castor oil may be attributed antisecretory effects. the values are mean ± SEM.95±0.514 25.52*** 61.25 cm.and stool demonstrates the efficacy of moringa castor oil induced enteropooling.29 mL.66 V HMO 500 46.055*** 28. The extract sufficiently the motility in charcoal meal test.50±1.35±0.59±2.001 vs control. centimeter. a decrease Further.99±3.86 V HMO 250 1.24±2. The Moringa oleifera leaves showed significant significant reduction in frequency of defecation.94 III Loperamide 3 0. Millilitres.10±1. +ve.04*** 55..7 22. The values are mean ± SEM. which acts primarily in the small All the doses of tested.103*** 57. Groups Treatment Dose mg/kg Mean volume of intestinal fluid (ml) ± SEM % Protection I -ve control 1 mL 1. These prostaglandins then cause an Discussion: increase of intestinal motility and secretions into The results of the present study could suggest the lumen of the intestine [22].54*** 48.20±1.20 85.30 26.01 and ***p<0. ***p<0.38*** 58. it is supported by the release of in the movement of charcoal from pylorus to prostaglandins.08*** 81.36 V1 HMO 500 1.86±2.54 sulfate III HMO 100 43.33±1. n=6. PGE2.2±1. inhibitory activity against castor oil and number of faecal droppings and mean weight of magnesium sulfate induced diarrhea. The extract also posse’s increase in the electrolyte secretion by creating an non-specific spasmolytic activities as it inhibited osmotic imbalance [23].12±2. lipases in the small bowel to glycerol and ricinoleic acid.3±1.079*** 72.04±0.87±2. μg. which results from the caecum was observed as given in Table 5.01±3.00 IV HMO 100 0.49±0.83±2. hyperperistalsis.067 -- II +ve control PGE2 100μg 2.490 27.66*** 44. n=6. al. **p<0.001 vs control. In a model of charcoal meal induced electrolytes and speed up the intestinal transit. Magnesium sulfate similarly causes an charcoal meal test in rats. Hydroalcoholic extract of to an anti-electrolyte permeability action.24 II Atropine 5 44. inflammation and irritation effect of the ricinoleic acid [21].40±3. Lakshminarayana et. Groups Treatment Dose Mean length Mean distance Mean % movement of % mg/kg of intestine travelled by charcoal ± SEM after Inhibition (cm)± SEM charcoal meal (cm) 30min ± SEM I Control 1mL 47.75*** 51. a significant oleifera leaves as an effective antidiarrhoeal reduction in gastrointestinal motility during agent.7±0.58±0. negative. -ve.10 87.1±0.35*** 38. the extract showed a intestine to stimulate secretion of fluid and significant protection against PGE2 and castor oil 72 .27 14. Table 5: Effect of hydroalcoholic extract of Moringa oleifera (HMO) on charcoal meal induced hyperperistalsis.. at all the doses tested. positive.25*** 41.50±1.35 IV HMO 250 46. microgram.80±1.02 40.

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