Biochemical and Radio Labeling Studies of Venom Naja naja karachiensis with its Neutralization by Medicinal Plants of Pakistan

Abstract

Biochemical and Radio Labeling Studies of Venom Naja naja karachiensis with its Neutralization by Medicinal Plants of Pakistan Background Snake bite envenomation is one of the vivid examples of neglected occupational hazards that accounts for tens of thousands of deaths all over the world. One of such instance is Naja naja karachisis bite, a nightmare for the inhibitants of Southern Punjab (Paksitan), often endup with countless deaths and sequela. To address this problem present study was designed to highlight scientific grounds for Naja naja karachisis envenomation and to rationalize folklore claimed Pakistani medicinal plants as a first aid treatment before proper hospitalization. Methods Proteomic characterization of Naja naja karchiensis venom was carried out with electrophoresis (SDS-PAGE) and HPLC (SEC & RP-HPLC) coupled LC-MS/MS whereas inorganic constituents were quantified with ICP-OES technique. Bio distribution and kinetic profile of venom was monitored with short lived radiotracer (99mTc) via direct radio isotopic binding technique. Lethal biological effects of crude venom were examined in terms of its LD50, hemolytic and anticoagulant behavior while toxic biochemical parameters (in vivo), towards liver (AST & ALT), heart (CK-MB & LDH) and kidneys (urea & creatinine) damage were investigated by following the recommendations of DGKC and IFCC methods. Venom was analyzed for different enzymatic activities (PLA2, ALPase, 5ʹ-ND, hyaluronidase and protease) by adopting conventional biochemical assays (in vitro). Twenty eight medicinal plants of Pakistan were extracted with methanol by simple maceration process and thereafter used to reverse deleterious actions of cobra venom. RP-HPLC coupled bioassay guided fractionation technique was used to characterize bioactive constituent/metabolite(s), responsible for anti-PLA2 activity in Bauhinia variegata L extract.