FIGURE 2.

Sorafenib inhibits HSC activation by triggering ferroptosis in vitro. (A) Cell viability of HSC‐T6 cells with different concentrations of sorafenib for 24 h was detected by CCK‐8. Cytotoxicity of AML‐12 cells was assayed by LDH Kit. Data were presented as the mean ±SD of 3 independent experiments. *p < 0.05, **p < 0.01. (B) Fer‐1 (1 μM), DFO (100 μM), ZVAD‐FMK (10 μM) or necrosulfonamide (0.5 μM) were exposed to HSC‐T6 cells with or without of sorafenib (10 μM), and cell viability was assayed by CCK‐8. Data were presented as the mean ± SD of 3 independent experiments. *p < 0.05, **p < 0.01. (C) Mitochondria in control and sorafenib‐treated groups were observed by transmission electron microscope (Scale bar: 200 nm). HSC‐T6 cells were treated with sorafenib (5, 10, 15 μM) for 24 h. (D) Iron release, MDA content and GSH expression in cell lysates were detected by kits. Intracellular ROS generation was detected with DCFH‐DA probe. Data were presented as the mean ± SD of 3 independent experiments. *p < 0.05, **p < 0.01. N.S. not significant. (E) Western blot analyses of SLC7A11 and GPX4 proteins were performed. Data were presented as the mean ± SD of 3 independent experiments. *p < 0.05, **p < 0.01. N.S. not significant. (F) Western blot analyses of α‐SMA, COL1α1 and fibronectin proteins were performed. Data were presented as the mean ± SD of 3 independent experiments. *p < 0.05, **p < 0.01. N.S. not significant