Acrylamide induces ferroptosis in HSC-T6 cells by causing antioxidant imbalance of the XCT-GSH-GPX4 signaling and mitochondrial dysfunction

Acrylamide (AA) is a heat-induced food contaminant, mainly metabolized by the liver. Increasing evidences have proved that ferroptosis is linked to the pathogenesis of liver disease. In the current study, the underlying mechanism of AA-induced rat hepatic stellate (HSC-T6) cells ferroptosis was investigated by detecting changes in iron levels, expressions of ferroptosis-related proteins and indicators of mitochondrial dysfunction. The results showed that AA treatment led to iron levels increased and expressions of long-chain acyl-CoA synthase 4 (ACSL4), cyclooxygenase 2 (COX2) and ferritin heavy chain 1 (FTH1) proteins in HSC-T6 cells were all altered. Treatment with the ferroptosis inhibitor ferrostatin-1 (Fer-1) markedly reversed the impact of AA, suggesting that AA induced ferroptosis in HSC-T6 cells. Mechanistically, AA induced the onset of ferroptosis by affecting XCT-GSH-GPX4 antioxidant signaling. Moreover, AA created a peroxidative environment for ferroptosis by inducing oxidative stress in HSC-T6 cells through mitochondrial dysfunction, as evidenced by increased mitochondrial ROS (mtROS) release, mitochondrial membrane potential (MMP) depolarization, and decreased mitochondrial ATP. Our results indicated that AA resulted in mitochondrial dysfunction and ferroptosis, and dysregulation of XCT-GSH-GPX4 antioxidant signaling was a key factor in AA-induced ferroptosis.