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Gastroprotective potentials of the ethanolic extract of Mukia maderaspatana against indomethacininduced gastric ulcer in rats ab

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G. Gomathy , D. Venkatesan & S. Palani a

Department of Biochemistry, Valliammal College for Women, Chennai, Tamil Nadu, India b

Research Centre, Manonmanium Sundaranar University, Tirunelveli627012, Tamil Nadu, India

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c

Department of Biotechnology, Arunai Engineering College, Tiruvannamalai, Tamil Nadu, India d

Department of Biotechnology, Anna Bioresearch Foundation, Arunai Engineering College, Tiruvannamalai, Tamil Nadu, India Published online: 04 Dec 2014.

To cite this article: G. Gomathy, D. Venkatesan & S. Palani (2014): Gastroprotective potentials of the ethanolic extract of Mukia maderaspatana against indomethacin-induced gastric ulcer in rats, Natural Product Research: Formerly Natural Product Letters, DOI: 10.1080/14786419.2014.986726 To link to this article: http://dx.doi.org/10.1080/14786419.2014.986726

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Natural Product Research, 2014 http://dx.doi.org/10.1080/14786419.2014.986726

SHORT COMMUNICATION Gastroprotective potentials of the ethanolic extract of Mukia maderaspatana against indomethacin-induced gastric ulcer in rats G. Gomathyab*, D. Venkatesanc and S. Palanid

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Department of Biochemistry, Valliammal College for Women, Chennai, Tamil Nadu, India; bResearch Centre, Manonmanium Sundaranar University, Tirunelveli 627012, Tamil Nadu, India; cDepartment of Biotechnology, Arunai Engineering College, Tiruvannamalai, Tamil Nadu, India; dDepartment of Biotechnology, Anna Bioresearch Foundation, Arunai Engineering College, Tiruvannamalai, Tamil Nadu, India (Received 9 June 2014; final version received 7 November 2014)

This study investigated the protective effects of the ethanolic extract of Mukia maderaspatana against indomethacin-induced gastric ulcer in rats. Gastric ulceration was induced by single intraperitoneal injection of indomethacin (30 mg/kg b.wt.). M. maderaspatana extract produced significant reduction in gastric mucosal lesions, malondialdehyde and serum tumour necrosis factor-a associated with a significant increase in gastric juice mucin content and gastric mucosal catalase, nitric oxide and prostaglandin E2 levels. The volume and acidity of the gastric juice decreased in pretreated rats. The plant extract was evaluated in the gastric juice of rats, untreated has showed near normal levels in pretreated rats. The M. maderaspatana was able to decrease acidity and increase the mucosal defence in the gastric area, therefore justifying its use as an antiulcerogenic agent. Ranitidine significantly increased pH value and decreased pepsin activity and gastric juice free and total acidity. The antiulcer effect was further confirmed histologically. Keywords: Mukia maderaspatana; malondialdehyde; gastric ulcer; prostaglandin; nitric oxide

1. Introduction Gastric ulcer is a major health hazard in terms of both morbidity and mortality (Chaturvedi et al. 2007). Gastric ulcer, the most common disorder of gastrointestinal tract has multifunctional causes in its pathophysiology (Higham et al. 2002). The aetiology of gastroduodenal ulcers is

*Corresponding author. Email: [email protected] q 2014 Taylor & Francis

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influenced by various aggressive and defensive factors such as acid-pepsin secretion, parietal cell, mucus secretion, blood flow, mucosal barrier, cellular regeneration and endogenous protective agents (prostaglandins and epidermal growth factors; Repetto & Llesuy 2002). Several pharmaceutical products have been employed for the treatment of gastroduodenal ulcer and peptic diseases, resulting in decreasing mortality and morbidity rates, but they are not completely effective and produce many adverse effects (Rates 2001). The leaves of Mukia maderaspatana find a prominent place in the Siddha and Ayurvedic systems of medicine. Folkloric traditional medicine claims that the leaves and tender shoots are useful as aperient, diuretic, stomachic, antipyretic, antiflatulent, antiasthmatic, antitussive, antihistaminic, antibronchitic and as an expectorant, in addition to its prescription against vertigo and biliousness (Chatterjee & Pakrashi 1997; Muthu et al. 2006; Khare 2007; Loganathan 2007).The leaf-tea is administered for the alleviation of jaundice (Jayaweera 1982) and mupatena tea, the extract of the leaves and bark, is reported to be a good decongestant and a very good remedy for cough, cold and flu (Prakash Rao 2007). In this article, we report the results of our attempt to determine the capacity of the leaf extract for gastroprotective activity induced by indomethacin. 2. Results and discussion 2.1. Effect of Mukia plant extracts on indomethacin-induced gastric lesions Ulcer index was significantly increased ( p , 0.01) in the indomethacin-treated group of animals (group II) compared with normal animals (group I), (Supplementary Figure S1 – online only). Treatment with ethanol extract of Mukia showed a significant reduction ( p , 0.05 and p , 0.01; groups IV and V) in ulcer index compared with the indomethacin-treated group (group II). However, plant extract alone (group VI) did not show any significant effect on ulcer index. 2.2. Effect of Mukia plant extracts on the gastric juice analysis Supplementary Figures S2– S4 (online only) show the effect of Mukia plant extract on the gastric juice analysis. Supplementary Figure S2 (online only) shows that the administration of indomethacin caused significant decrease in pH value associated with a significant increase in gastric juice free (Supplementary Figure S3 – online only) and total acidity (Supplementary Figure S4 – online only). Pretreatment with Mukia plant extract produced an insignificant change in pH value and free and total acidity as compared with the indomethacin group. Pretreatment with ranitidine (RAN) either alone or with Mukia plant extract produced a significant increase in pH value and significant decrease in free and total acidity when compared with the indomethacin group. Supplementary Figure S5 (online only) shows that the administration of indomethacin caused a significant increase in gastric juice pepsin activity associated with significant reduction in gastric juice mucin content. Pretreatment with Mukia plant extract produced an insignificant change in pepsin activity and a significant increase in mucin content as compared with the indomethacininduced group II. Pretreatment with RAN either alone or with Mukia plant extract significantly decreased the gastric juice pepsin activity (as compared with the indomethacin group). Meanwhile, pretreatment with RAN did not produce any significant change in the mucin content. Indomethacin caused a significant ( p , 0.01) increase in the levels of pepsin activity and mucin content (Supplementary Figures S5 and S6 – online only; group II) when compared with a normal control group (group I), Administration of ethanol extract of M. maderaspatana (groups III and IV) significantly ( p , 0.05, p , 0.01, respectively) decreased the pepsin and mucin levels, as compared with the indomethacin-induced group (group II). Administration of Mukia plant extract ensures that these levels are retrieved normally, significantly ( p , 0.05,

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p , 0.01) when compared with group II. Plant extract alone (group VI) did not show any significant effect on pepsin and mucin levels. In this study, indomethacin injection, a representative of nonsteroidal anti-inflammatory drugs (NSAIDs) family, caused a remarkably significant increase in ulcer index, gastric juice free and total acidity, and pepsin activity. The ulceration induced by indomethacin is attributed mainly to the various processes, including the generation of reactive oxygen species, initiation of lipid peroxidation, infiltration of the leucocytes, induction of the apoptosis and inhibition of prostaglandin synthesis (Bech et al. 2000). The use of NSAIDs is considered to be the major risk factor in gastric ulcers. The volume of acid present in gastric secretion, which encompasses HCl, mucus, pepsinogen, bicarbonates, intrinsic factor and protein, reflects acid volume. Oral administration of RAN significantly reduced the ulcer index, gastric juice free and total acidity, and pepsin activity. Gastric acid decimation by RAN is attributed to its ability to antagonise the binding of histamine to the H2 receptor on the parietal cells (Banji et al. 2010). Oral administration of Mukia plant extract produced significant decrease in ulcerative index, with insignificant changes in free and total gastric acid values and pepsin activity. Therefore, this result reinforced the absence of antisecretory activity of Mukia plant extract and possible strengthening of gastroprotective factors such as antioxidant elements in this extract. 2.3. Effect of Mukia plant extracts on gastric mucosal lipid peroxides malondialdehyde and catalase activity Administration of indomethacin significantly raised the gastric mucosal malondialdehyde (MDA) and catalase (CAT) concentration observed in the control group, as compared with group II. Interestingly, all the pretreatments which used to produce significant reduction in gastric mucosal MDA and CAT concentration as compared with the indomethacin group. Mukia plant extract treatment reduced the gastric mucosal MDA as well as CAT concentration. While RAN pretreatment reduced the gastric mucosal MDA concentration, administration of plant extract significantly augmented the decrease of gastric mucosal MDA and CAT concentration (Supplementary Figures S7 and S8 – online only). In this study, the decreased mucin secretion in the indomethacin-administered rats indicated reduced ability of the mucosal membrane to protect the mucosa from physical damage and back diffusion of hydrogen ions. Mucosal damage can be easily caused by the generation of exogenous and endogenous active oxygen and free radicals (Naito et al. 1995). Indomethacin is known to induce the reactive oxygen metabolites in animal models, which may contribute to mucosal injury (Chattopadhyay et al. 2006). This might lead to aggravated tissue damage during stomach ulceration (El-Missiry et al. 2001). Indomethacin-induced stomach ulceration was accompanied with a severe oxidative stress in gastric tissue, causing damage to key biomolecules such as lipids. RAN treatment significantly reverted the indomethacin-induced changes in MDA and CAT. RAN, an antisecretory drug, has often been reported to possess antioxidant and immunosuppressive actions, which may be responsible for its antiulcerogenic activity (Lapenna et al. 1994; Ardestani et al. 2004). 2.4. Effect of plant extracts on the gastric mucosal nitrites/nitrate content In indomethacin treated group, gastric mucosal nitrites/nitrate content was significantly reduced, RAN pretreatment group showed significantly altered the gastric mucosal nitrites/nitrate. Meanwhile, pretreatment with plant extract significantly increased gastric mucosal nitrites/ nitrate content (groups IV and V, Supplementary Figure S9 – online only). Nitric oxide (NO) is an endogenous defensive factor for gastric cells and exhibits gastroprotective properties against different types of aggressive agents (Samini et al. 2002).

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Prostaglandin, a key molecule that stimulates the complex array of ulcer-healing mechanisms, gets synthesised in mucosal cells by cyclooxygenase (COX) enzymes.

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2.5. Effect of Mukia plant extracts on the gastric mucosal prostaglandin E2 level The synthesis of mucosal prostaglandin E2 (PGE2) was markedly suppressed by indomethacin when compared with that in the normal rats (Supplementary Figure S10 – online only). However, the mucosal synthesis of PGE2 in the pretreated with plant extract increased significantly compared with that of the indomethacin group, which increased the PGE2 level marginally. Indomethacin causes ulcer mostly on the glandular (mucosal) part of the stomach (Nwafor et al. 1996) by inhibiting prostaglandin synthesis through the inhibition of the COX enzymes (Rainsford 1987). 2.6. Effect of Mukia plant extracts on serum level of proinflammatory cytokine tumour necrosis factor-a Serum level of pro-inflammatory cytokine tumour necrosis factor-a (TNF-a) was significantly increased ( p , 0.01) in the indomethacin-treated group of animals compared with the normal animals (Supplementary Figure S11 – online only). Treatment with the ethanol extract of the Mukia plant extract showed significant ( p , 0.05 and p , 0.01; groups III and IV) decrease in the concentrations of serum TNF-a compared with the indomethacin-treated group (group II). Plant extract alone (group VI) did not show any significant effect on serum TNF-a levels. TNF-a is a proinflammatory cytokine secreted by macrophages increased during ulcerative stress (Hamaguchi et al. 2001), it is a potent stimulator of neutrophil infiltration into the gastric mucosa (Wei et al. 2003) and inducible NO expression (Calatayud et al. 2001). In this study, indomethacin significantly increased serum TNF-a as compared with the control group. 2.7. Histopathological examination Histological examination of the gastric mucosal tissue showed sharply defined ulcer crater at the site of exposure to aspirin almost reaching the submucosal layer and a deep alteration of glandular epithelium. Damaged mucosal epithelium, leucocyte infiltration and ulcerated area covered with inflammatory exudates were observed in aspirin-induced rats. Treatment with M. maderaspatana extract showed absence of ulcer crater, clearance of the necrosis and maintenance of the mucosal layers along with normal glands (Supplementary Figure S12 – online only). 3. Conclusion In the present study, the ethanolic extract of M. maderaspatana plant possessed significant antiulcer properties, thus supports the traditional use M. maderaspatana plant in the treatment of gastrointestinal disorders. Mukia plant extract can protect from indomethacin-induced gastric ulceration due to its antioxidative, anti-inflammatory and mucin-enhancing properties. The mechanism of its gastroprotective activity may be attributed to the reduction in gastric mucosal lipid peroxidation (MDA) and serum TNF-a along with elevation in gastric juice mucin content and gastric mucosal CAT, NO and PGE2 levels. The protection rendered by the co-treatment of Mukia plant extract and RAN was found to be better than that of Mukia plant extract alone and RAN alone. Consequently, Mukia plant extract could be used together with RAN for the treatment of gastric ulcer.

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Supplementary material Experimental details relating to this paper are available online, alongside Figures S1 – S12.

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References Ardestani SK, Janlow MM, Kariminia A, Tavakoli Z. 2004. Effect of cimetidine and ranitidine on lipid profile and lipid peroxidation in g-irradiated mice. Acta Med Iran. 42:198–204. Banji D, Singh J, Banji OJ. 2010. Scrutinizing the aqueous extract of leaves of Pedalium murex for the antiulcer activity in rats. Pak J Pharm Sci. 23:295–299. Bech PL, Xavier R, Lu N, Nanda NN, Dinauer M, Podolsky DK, Seed B. 2000. Mechanisms of NSAID-induced gastrointestinal injury defined using mutant mice. Gastroenterology. 119:699–705. Calatayud S, Barrachina D, Esplugues JV. 2001. Nitric oxide: relation to integrity, injury and healing of the gastric mucosa. Microsc Res Tech. 53:325–335. Chatterjee A, Pakrashi SC. 1997. The treatise on indian medicinal plants. Vol. 5. New Delhi, India: National Institute of Science Communication and Information Resources, CSIR. Chattopadhyay I, Bandyopadhyay U, Biswas K, Maity P, Banerjee RK. 2006. Indomethacin inactivates gastric peroxidase to induce reactive oxygen-mediated gastric mucosal injury and curcumin protects it by preventing peroxidase inactivation and scavenging reactive oxygen. Free Radic Biol Med. 40:1397–1408. Chaturvedi A, Kumar MM, Bhawani G, Chaturvedi H, Kumar M, Goel KR. 2007. Effect of ethanolic extract of Eugenia Jambolana seeds on gastric ulceration and secretion in rats. Indian J Physiol Pharmacol. 51: 131 –140. El-Missiry MA, El-Sayed IH, Othman AI. 2001. Protection by metal complexes with SOD-mimetic activity against oxidative gastric injury induced by indomethacin and ethanol in rats. Ann Clin Biochem. 38:694–700. Hamaguchi M, Watanabe T, Higuchi K, Tominaga K, Fujiwara Y, Arkawa T. 2001. Mechanisms and roles of neutrophil infiltration in stress-induced gastric injury in rats. Dig Dis Sci. 46:2708–2715. Higham J, Kang JY, Majeed A. 2002. Recent trends in admissions and mortality due to peptic ulcer in England, increasing frequency of haemorrhage among older subjects. Gut. 50:460–464. Jayaweera DMA. 1982. Medicinal plants (indigenous and exotic) used in Ceylon. Vol. 4. Colombo: National Science Council of Sri Lanka. Khare CP. 2007. India medicinal plants – an illustrated dictionary. Berlin: Springer-Verlag. Lapenna D, De Gioia S, Mezzetti A, Grossi L, Festi D, Marzio L, Cuccurullo F. 1994. H2-receptor antagonists are scavengers of oxygen radicals. Eur J Clin Invest. 24:476–481. Loganathan N. 2007. Poorveegam Thoguppu Nool Kalangiam. Puducherry, India: Poorveegam Research Trust (in Tamil). Muthu C, Ayyanar M, Raja N, Ignacimuthu S. 2006. Medicinal plants used by traditional healers in Kancheepuram district of Tamil Nadu, India. J Ethnobiol Ethnomed. 2:43. doi:10.1186/1746-4269-2-43. Naito Y, Yoshikawa T, Matsuyama K, Yagi N, Arai M, Nakamura Y, Nishimura S, Yoshida N, Kondo M. 1995. Effects of oxygen radical scavengers on the quality of gastric ulcer healing in rats. J Clin Gastroenterol. 21:S82–S86. Nwafor PA, Effraim KD, Jacks TW. 1996. Gastroprotective effects of aqueous extracts of Khaya senegalensis bark on indomethacin-induced ulceration in rats. West Afr J Pharmacol Drug Res. 12:46–50. Prakash Rao EV. 2007. Herbal medicine as sustainable livelihood: a case of Irula Tribal Women Welfare Society from rural India. Asia Pac J Rural Dev. 17:85–100. Rainsford KDD. 1987. The effects of 5-lipoxygenase inhibitors and leukotriene antagonists on the development of gastric lesions induced by nonsteroidal anti-inflammatory drugs in mice. Agents Action. 21:316–319. Rates SMK. 2001. Plants as source of drugs. Toxicon. 39:603–613. Repetto MG, Llesuy SF. 2002. Antioxidant properties of natural compounds used in popular medicine for gastric ulcers. Braz J Med Biol Res. 35:523–534. Samini M, Moezi L, Jabarizadeh N, Tavakolifar B, Shafaroodi H, Dehpour A. 2002. Evidences for involvement of nitric oxide in the gastroprotective effect of bromocriptine and cyclosporin A on water immersion stress-induced gastric lesions. Pharmacol Res. 46:519–523. Wei XM, Heywood GJ, Di Girolamo N, Thomas PS. 2003. Nicorandil inhibits the release of TNF alpha from a lymphocyte cell line and peripheral blood lymphocytes. Int Immunopharmacol. 3:1581–1588.

Gastroprotective potentials of the ethanolic extract of Mukia maderaspatana against indomethacin-induced gastric ulcer in rats.

This study investigated the protective effects of the ethanolic extract of Mukia maderaspatana against indomethacin-induced gastric ulcer in rats. Gas...
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