Detection of Apoptosis Initiated in Treated HepG2 Cells with t-BHP: The Role of Phytochemicals to Reduce Toxicity and Stop Apoptosis

Abstract

Apoptosis or programmed cell death is a standard physiological mechanism. It is essential to control the number of cells, balance cell division and cell death, regulate the immune system, and eliminate pathogen-infected cells. Apoptosis entailed a different investigation to determine related biochemical reactions such as activated caspase, Reactive Oxygen Species (ROS), Lipid Peroxidation (LPO), and Evaluation of Glutathione Content (GSH) by using different techniques. HepG2 cells were exposed to +/- 0.4 and 0.8 mM t-BHP for specific times to induce toxicity for apoptosis detection. We aim to investigate the mechanism of cell death in treated HepG2 with t-BHP under consideration of the conditions of the cytoprotection assay. Results showed no strong evidence for apoptosis, although caspase-3 activity increased significantly (p ≤ 0.05) in treated HpG2 cells with 0.8 mM t-BHP at 150 minutes. The weak proof for apoptosis may attribute to the participation of Calpain through the cross-talk in blocking the caspase- activation. Similarly, we obtained significant ROS and lipid peroxidation increases in treated HepG2 cells with 0.8 mM t-BHP (p ≤ 0.05 and 0.01 respectively) at 150 minutes. Moreover, reported a (non-significant) decline in GSH amounts. Treatment of the cells with Q and I3C under the conditions used in the cytoprotection study prevented the weak activation of caspase-3 identified by western blot.