EVALUATION OF ANTI-DIARRHEAL AND DIURETIC ACTIVITY OF TABERNAEMONTANA DIVARICATA LINN. LEAVES

EVALUATION OF ANTI-DIARRHEAL AND DIURETIC ACTIVITY OF TABERNAEMONTANA DIVARICATA LINN. LEAVES

Sangita Kumari ^{* 1, 2}, A. Mazumder ¹ and S. Bhattacharya ³

Noida Institute of Engineering and Technology ¹, Institute of Pharmacy, Greater Noida - 201306, Uttar Pradesh, India.

Uttarakhand Technical University ², Dehradun - 248007, Uttarakhand, India.

Department of Pharmacy ³, College of Medicine and Health, Wachemo University, Ethiopia.

ABSTRACT: The aim of the present work was to investigate the anti-diarrheal and diuretic potentials of Tabernaemontana divaricata Linn. leaves of the family Apocynaceae. In almost all tropical developing countries of the world, diarrhoea is one of the most common and serious diseases, the principal cause of morbidity and mortality among children under five years. The methanolic extract of the leaves of T. divaricata L. was evaluated for anti-diarrheal properties in castor-oil induced diarrhoea. In the castor oil induced diarrhoea, MELTD significantly inhibited the number of defecations. The diuretic activity of the methanolic extract of leaves of T. divaricata (MELTD) was assessed based on Lipschitz value and diuretic index. The standard diuretic drug furosemide exhibited a significant increase in the urinary excretion of water and electrolytes (Na⁺ and K⁺), which is comparable with the extract. The diuretic index at the dose of 200 mg/kg (MELTD) was calculated as 1.55, significant as compare to the standard drug having a diuretic index of 1.70.

Anti-diarrheal, Diuretic, Tabernaemontana divaricata, MELTD, Defecations

INTRODUCTION: Diarrhoea is kenned as an incrementation in the number of stools (3 or more per 24 h), an incrementation in the fluidity of the stool and the presence of mucus or/and blood, with Neutrophils and polymorphs in the stool. Due to a severe loss of fluid and electrolytes in the stool, it leads to dehydration, hyponatremia, and hypokalaemia. Annually diarrheal diseases caused several million deaths in the world ¹. Diarrhea accounts for more than 8 million deaths worldwide each year in < age 5, especially in developing countries ^{2, 3}.

The impact of diarrheal infection is exacerbated by lack of adequate and affordable healthcare in developing countries, particularly in rural and marginalized areas ⁴. The WHO has initiated the Diarrhoea Disease Control Program, in which traditional medicinal practices and the evaluation of health education and prevention approaches were included ^{5, 6}.

These plants and their active constituents have been experimentally screened for their gastroprotective properties ⁷. Asian traditional medicine systems have identified several herbs and spices to treat GI tract disorders. There are many herbal plants which are listed in the indigenous system for their anti-diarrheal activity with lesser side effects. Diuretic drugs increase the rate of urine flow, sodium excretion and adjust the volume and composition of body fluids in a variety of clinical situations. Diuretic agents have very wide application in the treatment of various chronic diseases associated with edema. They are prescribed for the treatment of hypertension, congestive heart failure, nephritic syndrome glaucoma, diabetes insipidus, liver ailments and pregnancy toxemia ⁸.

The herbal plants used in traditional medicine as diuretics have increased in recent years and used in the treatment of hypertension. Natural diuretics increase the urine output and urinary electrolyte concentration ⁹. Naturally occurring diuretics inhibit Na⁺ re-absorption and also inhibit secretion of anti-diuretic hormones ¹⁰. The wide ranges of phytoconstituents were responsible for diuretic activity includes alkaloids, glycosides, tannins, phenolics coumarins and triterpenoids, etc. ^{11, 12}.

Tabernaemontana, commonly known as Tagar belongs to the family Apocynaceae is a beautifully shaped evergreen shrub distributed throughout India, Bangladesh and other parts of the South East Asia ^{13, 14}. The genus of Tabernaemontana includes approximately 100 species, commonly distributed in tropical countries of the world, including India, Brazil, Egypt, Sri Lanka, Vietnam, Malaysia and Thailand. Tabernaemontana divaricata (synonym- Ervatamia coronaria) commonly called as Crepe Jasmine, a glabrous, evergreen, dichotomously branched shrub mostly grown in tropical countries.

In traditional medicine, T. divaricata (L.) R.Br.is utilized in treatment of the diseases like diarrhea, abdominal tumors, arthralgia, asthma, epilepsy, ocular perceiver infections, pyrexia, fractures, headache, inflammation, leprosy, mania, oedema, paralysis, piles, rabies, rheumatic pain, skin diseases, urinary disorders, strangury, toothache, ulceration and regurgitating ¹⁵.

MATERIALS AND METHODS:

Plant Collection and Authentication: The aerial part of the Tabernaemontana divaricata was collected in the month of September from Greater Noida, Uttar Pradesh, India. The herbarium of the plant specimen Tabernaemontana divaricata (L.) R.Br.ex Roem and Schult. was authenticated by Dr. K.C. Bhatt, Senior Scientist, NBPGR, Indian Council of Agricultural Research, New Delhi with voucher number NHCP/NBPGR/2011-64/. The herbarium of this plant specimen is similarly preserved in the NIET institute.

Extraction: Tabernaemontana divarivata leaves were collected, dried in the shade and powdered. The dried powder was subjected to Soxhlet extraction using methanol at 40-60 ºC. The liquor was concentrated to recover methanol, and the extracts were kept in a desiccator for drying. The percentage yield was calculated as 10.8%. The dried methanol extracts of the leaf of T. divaricata (MELTD) was stored in a tightly closed amber color glass bottle.

Experimental Animals: Animal experiments were approved by the Institutional Animal Ethics Committee (IAEC) of Noida Institute of Engineering and Technology (Pharmacy Institute), Greater Noida (Reference no.1121acCPCSEA 7). Adult wistar rats (150-250 gm) of either sex were used for the study. The animals were maintained at the Animal House of the NIET Pharmacy Institute maintained under standard hygienic conditions. They were fed with standard diet and had free access to water ad libitum. The animals were maintained under standard conditions of humidity, temperature, and 12h light / 12h darkness cycle.

Anti-diarrheal Activity: Anti-diarrheal activity of methanolic extract of leaves of Tabernaemontana divaricata (MELTD) was assessed by castor oil induced diarrheal method.  The Wister rats of either sex (weight 150-200g) were divided randomly into groups with five animals in each group (n=5) and fasted for 18 h. Normal saline was administered to the control group, and the different doses of MELTD were administered in treated groups. Loperamide was used as standard drug.

One h later, all the animals were administered with 1 ml/rat of castor oil orally by gavage. The severity of diarrhoea was assessed at 1, 2, 4 h for 6 h. The total number of feces (both diarrheal and non-diarrheal) expelled was compared with the control group. The anti-diarrheal effect of the drug was calculated by considering the total number of diarrheal feces of the control group as 100% ¹⁶.

Procedure: The Wister rats were divided into fourteen groups with six animals in each group.

Group 1: Normal saline (5ml/kg p.o) and served as control.

Group 2: Standard drug, loperamide (5 mg/kg, orally)

Group 3, 4 and 5: MELTD (50, 100 and 200 mg/kg body weight, p.o), respectively.

The anti-diarrheal activity was expressed as % inhibition of diarrhoea. The percent (%) inhibition of defecation was measured using the following formula:

% Inhibition of defecation = A – B / A × 100

Where, A = mean number of defecation time caused by castor oil, B = mean number of defecation time caused by drug or extract.

Diuretic Activity: The diuretic activity was performed by the Lipschitz method, is based on the principle that water and sodium excretion in test animals is different as compared to rats treated with a high dose of urea. The “Lipschitz value” is the quotient between excretion by test animals and excretion by the urea control. Six adult Albino rats per group (150-250 gm) were kept in metabolic cages, provided with a funnel and wire mesh at the bottom to collect the urine. The urine excretion was recorded after 5 h. The sodium and potassium content was determined by flame photometer.

Procedure: The albino rats were divided into fourteen groups with six animals in each group.

Group 1: Normal saline (20 ml/kg p.o) and served as control.

Group 2: Standard drug, furosemide (20 mg/kg i.p).

Group 3, 4 and 5: MELTD (50, 100 and 200 mg/kg body weight, p.o), respectively.

The total urine volume was collected at the end of 5h. The parameters accounted for ascertaining the diuretic activity will be the total volume of urine and the urine concentration of Na⁺ and K⁺. Furthermore, two important indices such as diuretic index and Lipschitz value were determined using the following formula ^{16, 17}.

Diuretic index = Urinary excretion volume of the test group / Urinary excretion volume of the control group

Lipschitz value = Urinary excretion volume of the test group / Urinary excretion volume of the treated group

The Lipschitz value of 1.0 and more are regarded as a positive effect.

RESULTS AND DISCUSSION: Diarrhoea induced by castor oil is described as an appropriate model of the complex, prolonged processes of hypersecretion and accelerated transit that characterizes secretory diarrhea. The induction of diarrhea due to the action of ricinoleic acid formed by hydrolysis of the castor oil ^{18, 19}. Ricinoleic acid alters in the transportation of water and electrolytes resulting in a hypersecretory response ²⁰. Ricinoleic acid also sensitizes the intramural neurons of the gut. The neuronal sensitization promotes the removal of soft intestinal content further.

Evaluation of Anti-diarrheal Activity Using MELTD: The MELTD was evaluated for anti-diarrheal properties in castor oil-induced diarrhoea. Castor oil induces diarrhoea by increasing the volume of intestinal content by prevention of the re-absorption of water. The liberation of ricinoleic acid results in irritation and inflammation of the intestinal mucosa, leading to the release of prostaglandins, which results in stimulation of secretion. Diarrhoea was clinically apparent in all rats of the positive control group after 30 min of oral administration of castor oil for the next 4 h. The methanol extract of T. divaricata showed a marked anti-diarrheal effect in the rats as shown in Table 1.

TABLE 1: EFFECT OF MELTD ON CASTOR OIL INDUCED DIARRHOEA

The one way ANOVA and Dunnet’s Multiple comparison test was used for statistical analysis. Values are expressed as mean ± SEM (N=6), *P<0.05, **P<0.01, when compared with the control group.

The doses of 100 mg/kg and 200 mg/kg of MELTD extract produced significant (P<0.01) defecation. The doses of 100 mg/kg and 200 mg/kg decreases the 49.7% and 61.7% respectively of wet feces produced upon administration of castor oil, compared to the standard group administered loperamide (71.1%) at the dose of 5 mg/kg.

Evaluation of Diuretic potentials of MELTD: The determination of Lipschitz value was useful in the assessment of diuretic activity ²¹. The standard diuretic drug furosemide exhibited a significant increase in the urinary excretion of water and electrolytes (Na⁺ and K⁺), which is comparable with the extract. The Lipschitz value for MELTD at dose of 50 mg/kg, 100 mg/kg and 200 mg/kg were calculated as 0.64, 0.82 and 0.91 respectively.

As shown in Table 2, the urinary excretion of water and electrolytes exhibited by furosemide and MELTD doses were 70%, 25.8%, 40%, and 55.3% respectively, as compared to control group throughout 5 h.

TABLE 2: EFFECT OF MELTD ON DIURETIC ACTIVITY

The one way ANOVA and Dunnet’s Multiple Comparison Test was used for Statistical analysis. Values are expressed as mean ± S.E.M. (N=6), *P<0.05, **P<0.01, when compared with the control group.

CONCLUSION: Worldwide and especially in developing countries, diarrhoea is a major cause of deaths among the children below 5 years. The entire pharmaceutical industry is looking for the new chemical entity from a natural source, as natural agents are devoid of side effects or have very less side effects. The methanolic extract of T. divaricata leaves showed the promising result as an anti-diarrheal agent when compared to the standard moiety loperamide. The MELTD showed a significant decrease in several wet feces produced upon administration of castor oil as compared to the control group. Recently, researchers are focusing on safe and economic therapeutic agents. The use of herbal plants mentioned in traditional medicine as diuretics have increased recently in years. The MELTD showed the Lipschitz value and diuretic index of 0.91 and 1.55 respectively, indicating the MELTD can be beneficial in the development of new moiety having good diuretic potentials.

ACKNOWLEDGEMENT: Nil

CONFLICT OF INTEREST: Nil

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    How to cite this article:

    Kumari S, Mazumder A and Bhattacharya S: Evaluation of anti-diarrheal and diuretic activity of Tabernaemontana divaricata Linn. leaves. Int J Pharmacognosy 2018; 5(7): 409-13. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.5(7).409-13.

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