Figure 8.

NDUFA4L2 promoted lipid oxidation and ROS production through the p38/5‐LO pathways and increased HPASMCs proliferation by 5‐LO‐dependent lipid peroxidation. A, The levels of lipid peroxidation metabolites 4‐HNE and MDA were increased after overexpression of NDUFA4L2 in HPASMCs in vitro and reversed by p38 inhibitor SB203580 or si5‐LO; (B) DCFH‐DA for ROS detection showed the ROS amounts in different experimental groups; (C) Cell cycles were analysed the proliferation of HPASMCs after knockdown of 5‐LO in hypoxia condition. The histogram showed the percentage of cells in the G₀/G₁, S and G₂/M phases; (D) EdU staining was analysed the proliferation of 5‐LO‐dependent lipid peroxidation in regulation of HPASMCs proliferation in hypoxic condition; (E) CCK‐8 analysed the proliferation of HPASMCs under normoxia, hypoxia and hypoxia + si5‐LO; (F) Western blotting examined PCNA and cycle‐related protein expression after knockdown of 5‐LO in hypoxia condition; (G) The expression of p‐5‐LO/5‐LO was increased in overexpression of NDUFA4L2 in HPASMCs in vitro and reversed by 5‐LO silencing; (H) The protein expression of PCNA and cycle‐related proteins was determined by Western blot analysis. NC, negative control; Nor, normoxia; Hyp, hypoxia. Scale bar = 50 μm. ^* P < 0.05, ^** P < .01, ^*** P < .001. n = 6. All of the values are denoted as mean ± SEM