ANTI-ULCER STUDY OF STANDARDIZED ETHANOL ROOT EXTRACT OF AGANOSMA DICHOTOMA AND ISOLATED URSOLIC ACID
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 â‰¥ 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.
2. Groenen MJ, Kuipers EJ, Hansen BE, Ouwendijk RJ. The incidence of duodenal ulcers and gastric ulcers in a Western population: Back to where it started. Can J Gastroenterol 2009;23:604-8.
3. Miller TA. Mechanisms of stress-related mucosal damage. Am J Med 1987;83:8-14.
4. Kangwan N, Park JM, Kim EH, Hahm KB. The quality of healing of gastric ulcers: Natural products beyond acid suppression. World J Gastrointest Pathophysiol 2014;5:40-7.
5. Khan MSA, Jais AMM, Afreen A. Prostaglandin analogue and anti-oxidant activity mediated gastroprotective action of Tabernaemontana divaricata (L.) R. Br. Flower methanolic extract against chemically induced gastric ulcers in rats. Biomed Res Int 2013;2013:1-18.
6. Rao ChV, Ojha SK, Radhakrishnan K, Govindarajan R, Rastogi S, Mehrotra S, et al. Antiulcer activity of Utleria salicifolia rhizome extract. J Ethnopharmacol 2004;91:243-9.
7. Hewageegani HGSP, Ariyawansal HAS, Ratnasooriya WD. Gastroprotective activity of the paste of Evolvulus alsinoides L. (vishnukranti kalka) in rats. Vidyodaya J Sci 2006;13:23-31.
8. Alkofahi A, Atta AH. Pharmacological screening of the antiulcerogenic effects of some Jordanian medicinal plants in rats. J Ethnopharmacol 1999;67:341-5.
9. Schmeda-Hirschmann G, Yesilada E. Traditional medicine and gastroprotective crude drugs. J Ethnopharmacol 2005;100:61-6.
10. Wiart C. Ethnopharmacology of medicinal plants: asia and the pacific. Human Press Inc; 2006. p. 16.
11. Don G. Aganosma. In: A general history of the dichlamydeous plants. Vol. 4. London: CJG and F Rivington; 1837. p. 77.
12. Middleton David J. A revision of Aganosma (Blume) G. Don (Apocynaceae). Kew Bull 1996;51:455-82.
13. Kirtikar KR, Basu BD. Indian medicinal plants. dehradun, India: M/s Bishen Singh Mahendra Pal Singh; 1975. p. 1587.
14. Vaidya BD, Kashyap L. Materia Medica of Ayurveda. New Delhi, India: Concept Publishing Company; 2000.
15. Trease GE, Evans WC. Pharmacognosy. Fifteenth Ed. W. B. Saunders Company Ltd., London; 2002.
16. Makkar HPS. Quantification of tannins in tree foliage: A laboratory manual for the FAO/IAEA co-ordinated research project on â€˜Use of nuclear and related techniques to develop simple tannin assays for predicting and improving the safety and efficiency of feeding ruminants of tanniniferous tree foliageâ€™. Joint FAO/IAEA of Nuclear; Techniques in Food and Agriculture, Animal Production and Health Sub-programme. FAO/IAEA working document; Vienna; 2000. p. 3â€“5.
17. Kumaran A, Karunakaran J. In vitro antioxidant activities of methanol extracts of five Phyllanthus species from India. Food Sci Tech 2006;40:344-52.
18. Baccou JC, Lambert F, Sauvaire Y. Spectrophotometric method for the determination of total steroidal sapogenin. Analyst 1977;102:458-65.
19. Gupta M, Bisht D, Khatoon S, Srivastava S, Rawat AKS. Determination of ursolic acid a biomarker in different Swertia species through high-performance thin layer chromatography. Chinese Med 2011;2:121-4.
20. Organization for Economic Co-operation and Development. Guidelines for testing of chemicals. Guidance no. 425, Acute oral toxicity: up and down procedure (UDP). Available from: http://iccvam.niehs.nih.gov/Supp/Docs/Fed Docs/OECD/OECD tg425.pdf. [Last accessed on 03 Oct 2008]
21. Singh GK, Kumar V. Acute and subchronic toxicity study of a standardized extract of Fumaria indica in rodents. J Ethnopharmacol 2011;134:992-5.
22. Shay M, 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;5:43-61.
23. Shanti A, Radha R, Jaysree A. Antiulcer activity of newly formulated herbal capsule. Asian J Pharm Clin Res 2011;4:86-9.
24. Sanyal AK, Mitra PK, Goel RK. A modified method to estimate dissolved mucosubstances in gastric juice. Indian J Exp Biol 1983;21:78-80.
25. Laloo D, Prasad SK, Sairam K, Hemalatha S. Gastroprotective activity of ethanol root extract of Potentilla fulgens Wall ex Hook. J Ethnopharmacol 2013;146:505-14.
26. Lowry OH, Rosenborough NJ, Farr AL, Randal RJ. Protein measurement with folin phenol reagent. J Biol Chem 1951; 193:265-75.
27. Mukhopadhyay K, Bhattacharya D, Chakrabarti A, Goel RK, Sanyal AK. Effect of banana powder (Musa sapientum var. paradisiaca) on gastric mucosal shedding. J Ethnopharmacol 1987;21:11-9.
28. Reyes-Chilpa R, Baggio CH, Alavez-Solano D, Estrada-Muniz E, Kauffman FC, Sanchez RI, et al. Inhibition of H+K+-ATPase gastric activity by flavonoids, coumarins and xanthones isolated from Mexican medicinal plants. J Ethnopharmacol 2006;105:167-72.
29. Griswold B, Humoller FL, McIntyre AR. Inorganic phosphate and phosphate ester in tissue extracts. Anal Chem 1951;23:192-4.
30. Sairam K, Priyambada S, Aryya NC, Goel RK. Gastroduodenal ulcer protective activity of Asparagus racemosus: an experimental, biochemical and histological study. J Ethno-pharmacol 2003;86:1-10.
31. Morsy MA, Heeba GH, Abdelwahab SA, Rofaeil RR. Protective effects of nebivolol against cold restraint stress-induced gastric ulcer in rats: role of NO, HO-1, and COX-1, 2. Nitric Oxide 2012;27:117-22.
32. Rajasekaran S. Antiulcer activity of a different extract of Kigelia pinnata in experimental animals. Asian J Pharm Clin Res 2014;7:80-3.
33. Katayama S, Shionoya H, Ohtake S. A new method for extraction of extravasated dye in the skin and the influence of fasting stress on passive cutaneous anaphylaxis in guinea pigs and rats. Microbiol Immunol 1978;22:89-101.
34. Corne SJ, Morrissey SM, Woods RJ. Proceedings: a method for the quantitative estimation of gastric barrier mucus. J Physiol 1974;242:116-7.
35. Das D, Banerjee RK. Effect of stress on the antioxidant enzymes and gastric ulceration. Mol Cell Biochem 1993;125:115-25.
36. Kakkar P, Das B, Viswanathan PN. A modified spectro-photometric assay of superoxide dismutase. Indian J Biochem Biophys 1984;21:130-2.
37. Beers RF, Sizer IW. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 1952;195:133-40.
38. Sedlak J, Lindsay RH. Estimation of total protein bound and non-protein sulfhydril groups in tissues with Ellmanâ€™s reagent. Anal Biochem 1968;25:192-205.
39. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351-8.
40. Goel RK, Bhattacharya SK. Gastroduodenal mucosal defence and mucosal protective agents. Indian J Exp Biol 1991;29:701-14.
41. Baggio CH, Frietas CS, Otofuji GDM, Cipriani TR, Souza LM, Sassaki GL, et al. a Flavonoid-rich fraction of Maytenus ilicifolia Mart. ex reiss protects the gastric mucosa of rodents through inhibition of both H+ K+ ATPase activity and formation of nitric oxide. J Ethnopharmacol 2007;113:433-40.
42. Wallace JL. Mechanisms of protection and healing: current knowledge and future research. Am J Med 2001;110:19-23.
43. Ikasari ED, Utomo AB, Setyopuspito A, Adhityaasmara D, Setyowati H. Evaluation of antiulcer activity of mucoadhesive micro granules containing ranitidine hydrochloride in experimental rats. Int J Curr Pharm Res 2017;9:45-9.
44. Joshi A, Joshi VK, Pandey D, Hemalatha S. Systematic investigation of ethanolic extract from Leea macrophylla: Implications in wound healing. J Ethnopharmacol 2016;191:95-106.
45. Joshi A, Sengar N, Prasad SK, Goel RK, Singh A, Hemalatha S. Wound healing potential of the root extract of Albizzia lebbeck. Planta Med 2013;79:737-43.
46. Pandey D, Joshi A, Hemalatha S. Quality control standardization and in vitro antioxidant activity of Aganosma dichotoma K Schum root. Pharmacogn J 2015;7:74-82.
47. Liu J. Pharmacology of oleanolic acid and ursolic acid. J Ethnopharmacol 1995;49:57-68.
48. Navarrete A, Trejo-Miranda JL, Reyes-Trejo L. Principles of root bark of Hippocratea excels (Hippocrataceae) with gastroprotective activity. J Ethnopharmacol 2002;79:383-8.
49. Lee JH, Lee DU, Jeong CS. Gardenia jasminoides Ellis ethanol extract and its constituents reduce the risks of gastritis and reverse gastric lesions in rats. Food Chem Toxicol 2009;47:1127-31.
50. Sumbul S, Ahmad MA, Asif M, Akhtar M. Role of phenolic compounds in peptic ulcer: an overview. J Pharm Bioall Sci 2011;3:361-7.
51. Borrelli F, Izzo AA. The plant kingdom as a source of anti-ulcer remedies. Phytother Res 2000;14:581-91.