The Role of Reactive Oxygen Species in Crotalus atrox Venom‐induced Cell Death

Abstract

Venomous snake bites impact humans all around the world. Anti‐venom treatments mitigate systemic effects such as vascular hemorrhage, platelet aggregation inhibition, and the activation of inflammatory mediators. However, hemorrhagic snake venom also causes a loss of cellular adhesion to extracellular matrix components resulting in massive local tissue damage. To better understand the mechanism in which venom induces local tissue damage human embryonic kidney cells (HEKS) were grown on PEI then stimulated with 500μg/ml Crotalus atrox venom for 4 and 10 hours. Alamar Blue assays were used to measure cell viability and results suggest a 14.5±8.6% (p<0.05) and 58.95±10.7% (p<0.05) reduction in cell viability at 4 and 10 hours, respectively. Cells stimulated with 500μg/ml venom for 10 hours stained 97.6±2.2% (p<0.05) positive for Trypan blue, suggesting the venom reduces membrane integrity. HEK cells stimulated with 500μg/ml venom for 10 hours in the presence of 200 units PEG‐catalase increased viability by 44.1±5.1% (p>0.001) compared to cells stimulated with venom alone. 2′,7′‐Dichlorofluorescin‐diacetate (DCF‐DA) was used to quantify reactive oxygen species during venom stimulation. HEK cells stimulated with 50μg/ml Crotalus atrox venom for 4 hours returned 1.3×106±6.5×104 relative fluorescent units (RFU) compared to the 3.3×103±1.9×102 for non‐stimulated controls (p<0.05), a 433‐fold increase suggesting reactive oxygen species such as hydrogen peroxide play a critical role in Crotalus atrox venom induced cell death.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.