Cytotoxic Effect of Iranian Vipera Lebetina snake Venom on HUVEC Cells

Abstract

Objective: Envenomation by heamotoxic snakes constituted a critical health occurrence in the world. Bleeding is the most sever consequence following snake bite with viperid and crothalid snakes. It is believed that the degradation of vascular membrane caused hemorrhage; in contrast, some suggested that direct cytotoxicity has role in endothelial cell disturbances. This study was to evaluate the direct toxicity effect of V.Lebetina crude venom on Human Umbilical Vein Endothelial Cells (HUVECs). Methods: The effect of V. lebetina snake venom on growth inhibition of HUVECs was determined by MTT assay and neutral red uptake test. The integrity of cell membrane through LDH release was measured with the Cytotoxicity Detection Kit. Morphological changes of endothelial cells also were evaluated using a phase contrast microscope. Result: crude venom induced a concentration dependent inhibition on HUVECs with an IC50 value of 11.77 μg/ml after 24 h of incubation at MTT assay and 10.52 μg/ml at neutral red uptake test. In MTT assay, V. Lebetina crude venom showed a cytotoxic effect on endothelial cells which was confirmed by the effect observed with neutral red assay. Also, crude venom caused disturbances in the integrity of cell membrane by LDH release. The morphological alterations enhanced in high concentration and finally total cells number reduced. Conclusion: V. Lebetina venom showed potential direct cytotoxic effects on human endothelial cells in a manner of concentration dependent inhibition.