Chlorpyrifos induces cytotoxicity via oxidative stress and mitochondrial dysfunction in HepG2 cells

Chlorpyrifos (CPF), a widely used broad-spectrum organophosphate pesticide, has been associated with various adverse health effects in animals and humans. While its primary mechanism of action involves the irreversible inhibition of acetylcholinesterase, secondary mechanisms have also been suggested. The aim of the present study was to explore the secondary mechanisms of action involved in CPF-induced acute cytotoxicity using human hepatocarcinoma HepG2 cells. In particular, we investigated oxidative stress and mitochondrial function by assessing reactive oxygen species (ROS) generation, lipid peroxidation (LPO) and mitochondrial membrane potential (ΔΨ m ) alteration. Results showed that 24-h exposure to CPF (78.125–2500 μM) decreased cell viability in a concentration-dependent manner (IC 50 = 280.87 ± 26.63 μM). Sub-toxic CPF concentrations (17.5, 35 and 70 μM) induced increases in ROS generation (by 83%), mitochondrial superoxide (by 7.1%), LPO (by 11%), and decreased ΔΨ m (by 20%). CPF also upregulated Nrf2 protein expression, indicating the role of the latter in modulating the cellular response to oxidative insults. Overall, our findings suggest that CPF caused hepatotoxicity through oxidative stress and mitochondrial dysfunction. Given the re-emerging use of CPF, this study emphasizes the need for comprehensive analysis to elucidate its toxicity on non-target organs and associated mechanisms. • CPF induces acute cytotoxicity in human HepG2 cells (IC 50 = 280.87 μM). • Oxidative stress and mitochondrial disfunction were suggested as potential MoA. • Sub-toxic concentrations increase significantly ROS and mitoROS generation. • Overexpression of Nrf2 partially protects cells from CPF-induced oxidative stress. • CPF acute exposure significantly decreases mitochondrial membrane potential.