Publication Ref No.

: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-10/DEC/006

ISSN 0974 9446

EVALUATION OF ANTIULCER ACTIVITY POLYHERBAL FORMULATION Rakesh.A. Khandare*1, V.S. Gulecha1, M.S. Mahajan1, A. S. Mundada1, H.H.Gangurde1.
1

Mr. Rakesh Khandare

S.S.D.J. College of pharmacy, Neminagar, Chandwad (M.S.) India-423 101 Email: khandarera@gmail.com

ABSTRACT: The cause of ulceration in patients is mainly due to hypersecretion of gastric juice and also due to hypersecretion of pepsin. In traditional system of medicine a number of herbal preparations have been used for the treatment of peptic ulcers. The marketed polyherbal formulation (PHF) has been used for the treatment of gastrointestinal disorders. In view of this, in present study we have to evaluate the anti-ulcer activity of PHF. Study was carried out, by using two methods i.e., pyloric ligation and ethanol induced ulcers in rats pretreated with the doses of 1ml/kg Absolute ethanol, 100 mg/kg PHF, 50 mg/kg Ranitidine. After 4 hour in pyloric ligation method and 1hour in ethanol induced method rat was sacrificed and stomach was removed for observation of ulcer scores, ulcer index, free acidity, total acidity and pH. The PHF maintains the integrity of gastric mucosa by virtue of its effect on offensive and defensive gastric mucosal factors. PHF significantly (P<0.05) decreased free-acidity, total-acidity, ulcer index and gastric volume and significantly (P<0.05) increased the pH in pylorus ligated model whereas ulcer index significantly (P<0.05) decreased in ethanol induced model. Cyto-protective action may be the major mechanism responsible for the present study which cause the generation of prostaglandins promoting inhibition of ulcer. Our study shows that PHF has the potential to be used as an anti-ulcer.

KEYWORDS: Cyto-protective; Gastric ulcer; PHF;

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Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-10/DEC/006

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INTRODUCTION AND MATERIALS AND METHODS INTRODUCTION Peptic ulcer is an excoriated area of the gastric or duodenal mucosa caused by action of the gastric juice. It is a chronic and recurrent disease, and is the most predominant of the gastrointestinal diseases [1]. It is generally recognized that peptic ulcer is caused by a lack of equilibrium between the gastric aggressive factors and the mucosal defensive factors [2]. Gastric ulcer is among the most serious diseases in the world. The etiology of gastroduodenal ulcers is influenced by various aggressive and defensive factors such as acidpepsin secretion, parietal cell, mucosal barrier, mucus secretion, blood flow, cellular regeneration and endogenous protective agents such as prostaglandins and epidermic growth factors [3]. Some other factors, such as inadequate dietary habits, excessive ingestion of non-steroidal antiinflammatory agents, stress, hereditary predisposition and infection by Helicobacter pylori, may be responsible for the development of peptic ulcer [4]. In spite of the progress in conventional chemistry and pharmacology in producing effective drugs, the plant kingdom might provide a useful source of new antiulcer compounds for development as pharmaceutical entities or, alternatively, as simple dietary adjuncts to existing therapies [5]. The PHF is used for treatment of abdominal cramps and gastroprotective effect. The aim of present study was to evaluate the antiulcerogenic properties of marketed polyherbal formulation (PHF). Polyherbal formulation used in study contains Terminalia chebula, Emblica officinalis, Withania somnifera, Trigonella foenumgraecum, Glycerrhiza glabra, Piper nigrum, Bacopa monnieri, Zingiber officinale, Mucuna prurines, Asparagus racemosus, Ocimum sanctum and Curcuma longa. Some of these plants namely Withania somnifera, Bacopa monnieri, Asparagus racemosus, Ocimum sanctum, and Terminalia chebula have been classified in Ayurveda as rasayanas claim to improve mental and physical health [6-9]. Compound formulations are generally used in Ayurveda, based on the concept that such combination provides synergistic therapeutic effect. They also content ingredients, which are said to minimize the likely adverse effect of the major drug [10]. MATERIALS AND METHODS Ethanol induced ulcer The experiment was performed according to the method of Morimoto et al[12]. After 12 hour of fasting, the rats were randomly divided into three groups of six animals each. First group was given 1ml of vehicle (Distilled water), and the second group was treated with Ranitidine 50 mg/kg. The remaining groups received 100 mg/kg of PHF, respectively. All the treatments were administered orally. One hour after treatment, all the rats received 1ml of 99.5% ethanol to induce gastric ulcer. One hour later, the animals were sacrificed by cervical dislocation, and the stomachs removed and opened along the greater curvature. The stomachs were gently rinsed with water to remove the gastric contents and blood clots, for subsequent scanning. The ulcers were classified as 0 Normal stomach, 1- spot ulceration, 1.5- Hemorrhagic streaks, 2- Ulcer. Pyloric ligation: The assay was performed using the method of Shay et al [13] with a few modifications. The animals were divided into three groups (n = 6). After 24 h of fasting, the animals were anesthetized with thiopental sodium (10 mg/kg, i.p.), the abdomen was incised and the pylorus was ligated. Immediately after pylorus ligature, vehicle, PHF was administered dose of 100 mg/kg, Ranitidine 50 mg/kg to respective group. All International Journal of Pharma Research and Development Online 2

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Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-10/DEC/006 ISSN 0974 9446 the samples were administered orally. Four hours later, the animals were sacrificed by cervical dislocation; the abdomen was opened and another ligature was placed at the oesophageal end. The stomachs were removed and the gastric content collected and centrifuged at 3000 rpm (8000g, 250C, 10 min). The amount of gastric acid (ml) and the pH values were determined. The total acid secretion in the gastric hen was determined in the supernatant volume by titration to pH 7.0 using a 0.01M NaOH solution, and phenolphthalein as indicator.

Result: Absolute alcohol-induced ulcer model: In the ethanol-induced ulcer model, administration of ethanol produced haemorrhagic gastric lesions in the gastric mucosa of the control group. It was observed that the treatment with PHF 100 mg/kg and Ranitidine 50 mg/kg significantly (p < 0.05) reduced the lesion index, the total lesion area and the percentage of lesion, in comparison with negative control group (Fig 1). Pyloric ligation: The rat pretreated with PHF(100mg/kg) produced significant (P<0.01) decrease in ulcer index, gastric volume (Fig 2). As well as PHF also significantly (P<0.01) reduces free acidity, total acidity. Where as, pH was significantly (P<0.01) increased when campared with control group. Ranitidine also show smilier effects but was more effective compared with PHF (Fig 3). Discussion: There are several factors that may induce ulcer in human being such as stress, chronic use of antiinflammatory drugs and continuous alcohol ingestion, among others [15]. Although in most cases the etiology of ulcer is unknown, it is generally accepted that it is the result of an imbalance between aggressive factors and maintenance of the mucosal integrity through the endogenous defense mechanism [16, 17 ]. The candidate for an effective drug against peptic ulcer should basically act either by reducing the aggressive factors on gastroduodenal mucosa or by increasing mucosal resistance against them. It has become imperative to scrutinize herbal products for evaluating their acclaimed properties, as recently numbers of herbs are being introduced in the market. Keeping this view, we have attempted to study the PHF for its antiulcer activity by using different experimental models of gastric ulcer, pyloric ligation and ethanol induced ulcer, which operate by distinct mechanisms of ulcerogenesis. Oral administration of the damaging agent to the control group clearly produced a mucosal damage characterized by multiple hemorrhage red bands of different sizes along the long axis of the glandular stomach as described in other studies [13, 18]. Pretreatment with PHF (100 mg/kg) produced significant decrease in the intensity of gastric mucosal damages induced by the necrotizing agent ethanol compared with control group. In ethanol induced ulcer, the ulcer scores were significantly (P<0.05) decreased in rats pretreated with PHF which were compared with control group. Moreover the result indicates that; in case of pyloric ligation method, the ulcer index, free acidity, total acidity, and volume of gastric juice was significantly(P<0.05) reduced in rats pretreated with PHF where as pH was found to be significantly(P<0.05) increased in rats when compared with control group. Cyto-protective action by drugs has been considered to be due to the generation of prostaglandins or blockade of back diffusion of H+ ions [19] will be the major mechanism which is responsible for anti-ulcer activity. The PHF significantly reduced the gastric acid secretion in the present study. International Journal of Pharma Research and Development Online 3

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The cyto-protective action which promotes the generation of prostaglandin and causes decreases in secretion of gastric acid, mixture significantly reduced the gastric ulceration in pyloric ligated rats without affecting the gastric secretion or pepsin. But in case of ethanol induced method, the cyto-protective action has been decreases by ethanol, which is due to inhibition of synthesis of endogenous prostaglandin, which promote the formation of ulcer, the protective effect of PHF against ethanol induced ulcer in rats may show the anti-ulcer activity by decreasing the ulcer scores. The result further point out that, the cyto-protection may be major mechanism responsible for the antiulcer activity of the PHF. PHF may show the anti-ulcer activity against both models by generating the prostaglandin which causes inhibition of secretion of gastric fluid. Also it contains Terminalia chebula, which having property of protein precipitation and vasoconstrictor effect which could be advantages in preventing ulcer development [20]. The PHF contains Withania somnifera, Glycerrhiza glabra, Bacopa monnieri, Ocimum sanctum which is having antioxidant activity. As PHF contains more herbs which will be acting synergistically for producing antiulcer effect. The mechanism of action responsible for antinuclear activity of PHF may be cytoprotective action or antioxidant property of various herbs it contains. Further study required for finding out its exact mechanism of action which is underway. Reference: 1. Guyton and Hall. Textbook of Medical Physiology 10th Edn. Philadelphia 2000: p. 397-398. 2. Falcao HS, Mariath IR, Diniz MFFM, Batista LM, Barbosa-Filho JM. Plants of the American continent with antiulcer activity. Phytomed 2008 15: 132146. 3. Repetto MG, Llesuy SF. Antioxidant properties of natural compounds used in popular medicine for gastric ulcers. Braz J Med Biol Res 2002 35: 523534. 4. Peckenpaugh NJ, Poleman CM. Nutricao: Essencia Dietoterapia, 7th ed. Roca, Sao Paulo 1997. 5. Nadkarni AK, Indian Materia Medica, Vol-2 , Popular Prakashan, Page No. 245-270. 6. Wagner H, Norr H, Winterhoff H. Plant Adaptogens. Phytomed 1994 1:63-76. 7. Joshi H, Parle M. Zingiber officinale: Evaluation of its nootropic effect in mice. Afr J Trad CAM 2006 3: 64-74. 8. Joshi H, Parle M. Evaluation of nootropic potential of Ocimum sanctum Linn in mice. Ind J Expt Biol 2007 44: 133-136. 9. Kasture SB, Kasture VS, Joshua AJ, Damodarn A, Amit A. Nootropic activity of BacoMind TM , an enrinched phytochemical composition from Bacopa monnieri. J Nat Rem 2007 7: 166-173. 10. Bhattacharya SK. Nootropic effect of Mentat, a psychotropic formulation, on cognitive deficits induced by prenatal undernutrition, postnatal environmental impoverishment and hypoxia in rats. Indian J Exp Biol 1994. 32:31 36. 11. Quartini A, Negri E, Vecchia C La. Trends in peptic ulcer mortality in Italy, 1955-1985. J Epid Comm H 1992 46(5): 494 - 497. 12. Morimoto Y, Shimohara K, Oshima S, Sukamoto T. Effects of the new anti-ulcer agent KB-5492 on experimental gastric mucosal lesions and gastric mucosal defensive factors, as compared to those of teprenone and cimetidine. Jpn J Pharmacol 1991 57: 495-505. 13. Shay H, Komarov SA, Fels SS, Meranze D, Gruenstein M, Siplet H. A Simple Method for the Uniform Production of Gastric Ulceration in the Rat Gastroenterol 1945: 5:43. International Journal of Pharma Research and Development Online 4

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Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-10/DEC/006 ISSN 0974 9446 14. Nwafor PA, Okwuasaba FK, Binda LG. Antidiarrhoeal and antiulcerogenic effects of methanolic extract of Asparagus pubescens root in rats. J Ethnopharmacol 2000 72: 421 - 427. 15. Barocelli E, Chiavarini M, Ballabeni V, Barlocco D. Vianello P, Dal Piaz V, Impicciatore M. Study of the antisecretory and antiulcer mechanism of new indenopyridazinone derivative in rats. Pharmacol Res 1997 35: 487 - 492. 16. Piper DW, Stiel DD. Pathogenesis of chronic peptic ulcer, current thinking and clinical implications. Medical Progress 1986 2: 7 - 10. 17. Sergio FA, Marivane L, Eros C, Vania FN, Valdir CF, Rivaldo N. Evaluation of the antiulcerogenic activity of Maytenus robusta (Celastraceae) in different experimental ulcer models. J Ethnopharmacol 2007 113: 252257. 18. Yassir, M., Mulla A, HummadiYM, Najim, RA, Farjou IB. A new in vitro model for ethanol-induced gastric mucosal damage. Jap J Pharmacol Toxicol Meth 1999 41:167 - 172. 19. De B, Maiti RN, Joshi VK, Agrawal VK Goel RK. Effect of some Sitavirya drugs on gastric secretion and ulceration. Ind J Expt Biol 1997 35:1084 - 1087. 20. Tripathi KD. Essential of medical Pharmacy, Jaypee brothers medical publication, 4th edition 1994:627-639

TABLES AND FIGURES

Ulcer index

0 Ethanol Ranitidine 50 Treatment (mg/kg) PHF 100

Figure 1: Effect of PHF ulcer index in ethnol induced ulcer model.

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Publication Ref No.: IJPRD/2009/PUB/ARTI/VOV-1/ISSUE-10/DEC/006

9 8 7 6 5 4 3 2 1 0

ISSN 0974 9446

* * ** **

Vol of gastric secretion Ulcer Index

Ethanol

Ranitidine 50 Treatment (mg/kg)

PHF 100

Figure 2: Effect of PHF on ulcer index and volume of gastric secretion in pylorus ligation induced ulcer model.

70 60 50 40 30 20 10 0 pH

** ** *
Ethanol PHF 100 Treatment (mg/kg)

Free acidity Total acidity

*
Ranitidine 50

Figure 3: Effect of PHF on pH, Free acidity and total acidty in pylorus ligation induced ulcer model.

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