Original Article ANTI-ULCER STUDY OF STANDARDIZED ETHANOL ROOT EXTRACT OF AGANOSMA DICHOTOMA AND ISOLATED URSOLIC ACID DEEPALI PANDEY, APURVA JOSHI, SIVA HEMALATHA * Department of Pharmaceutics, Indian Institute of Technology (Banaras Hindu University), Varanasi 221005, India Email: shemalatha.phe@itbhu.ac.in Received: 06 Jan 2017 Revised and Accepted: 27 Feb 2017 ABSTRACT Objective: Aganosma dichotoma K. Schum (Apocynaceae) has been traditionally used as an Ayurvedic ulcer treatment and the study scientifically validates the antiulcer effect of A. dichotoma ethanol root extract (EAD). Methods: The studies included the isolation, quantification of ursolic acid through HPTLC. Acute and sub-acute toxicity study of EAD for 28 d and antiulcer effect of ursolic acid (50 mg/kg, p. o.), EAD (100, 200 and 400 mg/kg, p. o.) were also evaluated on both the physical (pyloric ligation, PL; cold restrain stress, CRS;) and chemical (absolute ethanol, aspirin, ASP;) induced ulcer in Charles Foster albino rats for 7 d. The various gastric-ulcer parameters, viz. gastric pH, volume, acid-pepsin output, DNA content, H + K + -ATPase activity, mucus content, microvascular permeability, antioxidant enzyme, and gastric histopathological study were performed. Results: The isolated ursolic acid was characterized by NMR and mass spectrometer and quantified through HPTLC in EAD (4.26% w/w). Acute oral toxicity study indicated that LD50 of extract was ≥ 5 g/kg. EAD at the dose of 200 and 400 mg/kg, p. o. reduced the ulcer score in both physical and chemical-induced ulcer models. In PL model EAD (400 mg/kg, p. o.) and ursolic acid (50 mg/kg, p. o.) showed antisecretory property by inhibiting aggressive factors [increase in gastric pH (35.02%, 26.73%), whereas decreased gastric volume (43.55%, 34.35%) and acid-pepsin output (75.23%, 68.81%), respectively]. EAD at 400 mg/kg p. o. showed significant effect on proton pump inhibition while ursolic acid didn’t showed any effect. Conclusion: The effects of EAD were accredited mainly to the offensive mechanism and justify its traditional usage in the treatment of gastric ulcers. Keywords: HPTLC, Gastric ulcer, Antisecretory, H + K + -ATPase inhibitor © 2017 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) DOI: http://dx.doi.org/10.22159/ijpps.2017v9i4.16957 INTRODUCTION Gastric ulcer is a widespread global problem in the modern era; it is estimated that around 14.5 million of the worldwide population is affected, with a mortality of 4.08 million [1]. The pervasiveness of gastric ulcer is due to behavioral and environmental factors such as inadequate dietary habits, smoking, excessive use of non-steroidal anti-inflammatory agents (NSAIDs), stress, hereditary pre- disposition, and infection with Helicobacter pylori [2,3]. Out of the above mentioned factors infection with H. pylori and long-term use of NSAIDs, are the most prevalent for the development of gastric ulcer [4]. Inhibition of gastric acid secretion by histamine receptor blockers, proton pumps inhibitors or enhancing the mucosal production (prostaglandin analogues or certain antimicrobials) are common strategies in modern ulcer treatment [5]. These available antiulcer drugs are associated with adverse complications including, hypersensitivity, arrhythmia, impotence, gynecomastia, and hematopoietic disorder [6, 7]. Currently, ethno-medicinal plants contribute to the drug development process and are a major focus in global health care. Plant extracts are important sources of new drug development and have been shown to produce potent results for the treatment of gastric ulcer [8, 9]. About 80 species of plants of the Apocynaceae family show medicinal importance and are often used to treat gastrointestinal ailments, reduce fever and pain, treat diabetes and infectious disease, etc. [10]. Aganosma dichotoma K. Schum (Apocynaceae) is commonly known as Malati in Hindi and Jati in Sanskrit. Eight species of Aganosma are found and distributed in India, China, Philippines, and Indonesia. In India, it mainly extends throughout Assam, Bihar, West Bengal, Orissa, Andhra Pradesh, and Tamil Nadu. Blume was first to describe Aganosma genus under class of Echites which attained the generic status by G. Don (1837) with other species based on specimens in Wallich's herbarium [11, 12]. A. dichotoma is traditionally used as anemetic, anthelmintic, in bronchitis, leprosy, skin diseases, ulcer, inflammations, arthritis, purulent discharges from the ear, and diseases of the mouth. Flowers are useful in eye diseases and leaves cure biliousness [13, 14]. Thus, the present investigation was aimed to establish the scientific validation of A. dichotoma in treatment of gastric ulcer. MATERIALS AND METHODS Materials Chemicals Tris buffer, Evans blue dye, Alcian blue dye, Topfers reagent and Folin-ciocalteu reagent were purchased from Sigma Aldrich (Germany). Some markers of analytical standards like Ursolic acid, Tannic acid, Rutin and Diosgenin were also purchased from Sigma Aldrich (Germany). Standard drug Omeprazole was obtained from Cadilla Pharmaceuticals Ltd. (India). All other chemicals and solvents used were of analytical grade. Plant material The roots of A. dichotoma were collected in the month of April 2013 from Tumbura Kona Kshetram at Seshachalam hills and Tirumala hills, Chittor District (13.2218 ° N, 79.1010 ° E), Andhra Pradesh, South India and authenticated by Dr. K. Madhava Chetty, Taxonomist, S. V. University, Tirupati. A voucher specimen (COG/AD/17) has been kept in Department of Pharmaceutics, IIT- BHU, Varanasi, India for further reference. Animals Adult Charles Foster albino rats (150±10 g) of either sex were obtained from the Central Animal House, Institute of Medical Sciences, Banaras Hindu University and were used for the study. The pharmacological experiments were performed after the approval obtained from the Central Animal Ethical Committee, Institute of International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 9, Issue 4, 2017