Effects of Aqueous Stem Extract of Achyranthes aspera on Bitis arietans Venom Protease and Phospholipase A2 Activity

American Journal of BioScience Volume 5, Issue 3, May 2017, Pages: 54-58 Received: Feb. 16, 2017; Accepted: Mar. 27, 2017; Published: May 27, 2017 Authors Hope Chinyere Nwune, Department of Biochemistry, Faculty of Science, University of Maiduguri, Maiduguri, Nigeria Mohammed Adamu Milala, Department of Biochemistry, Faculty of Science, University of Maiduguri, Maiduguri, Nigeria Hassan Zanna, Department of Biochemistry, Faculty of Science, University of Maiduguri, Maiduguri, Nigeria Abstract PDF (197KB) Aqueous stem extract of Achyranthes aspera was investigated for inhibitory activity against Bitis arietans venom protease and phospholipase A2 activity. The elemental analysis and phytochemical screening of the plant extract were carried out. The activities of protease and phospholipase A2 (Vo) of the crude Bitis arietans venom was determined and the data obtained was used to estimate KM, Vmax and Kcat. Inhibition studies were carried out using the same procedure except that different concentrations of the extracts (5%, 10%, 15% for protease assay and 0.5%, 0.75%, 10%, 1.25% and 1.5% for phospholipase A2 assay) were added to the reaction mixture. The result showed that the Bitis arietans venom protease had a Vmax of 0.062 ± 0.013 µmol/min, KM of 0.496 ± 0.095mg/ml and a Kcat of 0.125 ± 0.001min-1. The result also indicates that the Bitis arietans phospholipase A2 had a Vmax of 3.27 ± 0.030min-1, KM of 8.358 ± 0.050 mg/ml and Kcat of 0.391 ± 0.002min-. The aqueous stem extract produced a statistically significant (P<0.05) decrease in the Vmax, KM and Kcat of the Bitis arietans venom phospholipase A2 in a dose dependent manner and a statistically significant (P<0.05) increase in the Vmax, KM and Kcat of Bitis arietans protease in a dose dependent manner. The phytochemical screening revealed the presence of flavonoids, tannins, steroids, saponins and terpenoids in the extract while the elemental analysis revealed the presence of Zn, Cr, Ni, Cd, Mn, Fe and Na. The result suggests that aqueous stem extract of Achyranthes aspera inhibited the Bitis arietans venom phospholipase A2 in an uncompetitive manner while the protease activity was stimulated by the extracts. It was observed that the use of the stem of Achyranthes aspera may be important in the treatment of snake bites. Keywords Achyranthes aspera, Bitis arietans, Antivenom, Protease, Phospholipase A2 To cite this article Hope Chinyere Nwune, Mohammed Adamu Milala, Hassan Zanna, Effects of Aqueous Stem Extract of Achyranthes aspera on Bitis arietans Venom Protease and Phospholipase A2 Activity, American Journal of BioScience. Vol. 5, No. 3, 2017, pp. 54-58. doi: 10.11648/j.ajbio.20170503.13 Copyright Copyright © 2017 Authors retain the copyright of this article. This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. References [1]Abubakar, M. S., Abdulrahaman, E. M., Haruna, A. K. and Jahun, B. M. (2000). Invitro snake venom detoxifying action of leaf extract of Guierra senegalensis. Journal of Ethnopharmacology, 69: 253-257. [2]Aguiyi, J. C. (2011). The Molecular Basis of Natural Products Development, Unijos Inaugural Lecture Series 49, Journal of Natural, Knowledge and Health, Pp. 1-4. [3]Alam, M. I., Auddy, B. and Gomes, A. (1996). Viper venom neutralization by Indian medicinal plants (Hemidesmus indicus and Pluchea indica) root extract, Phytotheraphy Resources: 10: 58-61. [4]AOAC. Officia methods of analysis. Association of analytical chemist 15th ed. Washington DC: American Chemical Society. 1990; pp12-13. [5]Bradford, M. M. A. (1976). Rapid and Sensitive Method for the Qualification of Microgram Quantities of Protein Utilizing the Principle of Protein dye Binding. Journal of Analytical Biochemistry, 72: 248-254. [6]Dey, A. (2011). Achyranthes aspera L: Phytochemical and Pharmacological Aspects, international Journal of pharmaceutical sciences Review and research, 9 (2): 72-76. [7]Evans, W. C. and Sannders W. B. (2002). Plants in African Traditional Medicine – an Overview in Trease and Evans Pharmacognosy, 15th Edition, Pp. 488-495. [8]Fahmey, A. S., Ali, A. A. and Mohammed, S. C. (2004). Characterisation of Cysteine Protease from Wheat TriticumAestivum, Biores Technology, 91: 297. [9]Gomes, A., Das, R., Sarkhel, S., Mishara, R., Mukherjee, S., Bhattacharya, S. and Gomes, A. (2010). Herbs and herbal constituents active against snake bite. Indian Journal of experinmental biology, 48: 865-878. [10]Gray, J. E. (1842). Monographic synopsis of the vipers, or the family viperidae. Zoological Miscellany London, 2 (69): 68-71. [11]Harbone, J. B. (1973). Phytochemical Methods: A Guide to Modern Techniques of Plant Analysis. Chapman A and Hall London Pp. 279-280. [12]Ibrahim, M. A., Aliyu, A. B., Abusufiyanu, A., Bashir, M. and Sallau, A. B. (2011). Inhibition of Najanigricolis (Reinhard) Venom Protease Biology, 49: 552-554. [13]Jabeen, S., Muhammad, T. S., Khan. S. and Hayat, M. Q. (2010). Determination of major and trace elements in ten important folk and therapeutic plants of haripur basin, pakistan. Journal of medicinal plant research, 4 (7): 559-566. [14]Jager A. K (2015) Plant –based treatment of snakebites indian journal of traditional knowledge 14 (4): 571-573. [15]Lans, C., Harper, T., Georges, K., Bridgewater, E. (2001). Medicinal and ethnovetrinary remedies of hunters in trinidad BMC compliment. Alternative Medicine, 1: 1-10. [16]Meendatchisundaram, S., Parameswari, G. and Michael, A. (2009). Studies on the Antivenom Activity of AndrographisPaniculata and AristolichiaIndica Plant Extracts against Daboiaresseli Venom by Invivo and Iinvitro Methods, Indian Journal of Therapeutics Africa, 2 (4): 76-79. [17]Okonogi, Hatton T. Z., Ogiso, A. and Mitsui, S. (1979). Detoxification by PermisionTanin of Snake Venom and Bacterial Toxins. Toxicon Journal, 17: 524-527. [18]Rao, V. Y., Chakrabarti, R. (2004). Enhanced anti-proteases in Labeo Rohita fed with diet containing herbal ingredients. Indian journal of clinical biochemistry, 19 (2): 132-134. [19]Richards, A. D., Phylip, L. H., Farmerie, W. G., Scarborough, P. E., Alvarez, A., Dunn, B. M., Hirel, P. H., Strop, P. Pavilickova, L., Kostka, V., and Kay, J. (1990). Journal of Biochemistry, 265 (14). 7733. [20]Samy, R. P., Thwin, M. M., Gopalakrishnakone P. and Ignacimuthu S. (2008). Ethnobotaical survey of folk plants for the treatment of snake bites in southern part of Tamilnadu, Indian journal of ethnophamacology, 2: 39-45. [21]Sallau, A. B., Njoku, G. C., Olabisi, A. R., Wuruchekke, A., Abdulkadir, A. A., Isah, S., Abubakar, M. S. and Ibrahim, S. (2005). Effect of Guerra senegalensis leaf extract on some Echis carinatus venom enzymes. Journal of medical sciences, 5: 280-283. [22]Shendkar, C. D., Chandrachood, P. S., Pawar, A. B., Lavate, S. M. and Nirmala, R. and Deshpande, N. R. (2011). Quantitative estimation of macro, micronutrients and trace elements by X-ray fluorescence spectroscopy (XRF) from Achyranthes asspera International Journal of chemtech research, 3 (2): 610-613. [23]Sofowora, A. (1993). Medicinal Plants and Traditional Medicinal in Africa 2nd Edition. Sunshine House, Ibadan, Nigeria. Spectrum Books limited: Screening Plants for Bioactive Agents, Pp. 134-156. [24]Trease, G. E. and Evans, W. C. (2002). Pharmacognosy 15th edition, Saunder publishers Pp. 42-393. [25]Vrbka, L., Vondrasek, J., Jagoda-Cwiklik, B., Vacha, R., Jung, B. and Wirth P. (2006). Proceedings of the National academy of Sciences of the United States of America, 103 (42): 15440.