Comparative Study on Effects of Hypoxia Induced by Cobalt Chloride and Gas Hypoxia on Proliferation of Rat Primary Pulmonary Artery Smooth Muscle Cells
HU Yuedi1, ZHU Jie1,2,3*, ZHANG Lu2, WANG Xiaole2, TONG Jiabing2,3,4, CHENG Weiye1, LI Zegeng2,3
The purpose of this study was to compare the effects of CoCl2 (cobalt chloride) and gas hypoxia on proliferation and oxidative stress of rat primary PASMCs (pulmonary artery smooth muscle cells). PASMCs were stimulated by different concentrations of CoCl2 (25 μmol/L, 50 μmol/L, 100 μmol/L, 200 μmol/L, 400 μmol/L), or were placed in a hypoxic incubator. The effects of CoCl2 and gas hypoxia on the proliferation of PASMCs were compared by MTT assay, CCK-8 assay, and EdU assay. Under CoCl2 or gas hypoxia, the expression of hypoxic sensitive protein HIF-1α (hypoxia inducible factor-1α) and intracellular ROS (reactive oxygen species) were detected by Western blot and Flow cytometry. The results showed that the cell proliferation, expression of HIF-1α and ROS increased in PASMCs stimulated by CoCl2 or gas hypoxia for 24 h. Compared with low (25 μmol/L), medium (200 μmol/L), and high (400 μmol/L) concentrations of CoCl2 groups, the cell proliferation, the expression of HIF-1α protein, and intracellular ROS content were markedly augmented under gas hypoxia. Compared with low and high concentrations of CoCl2 groups, medium concentration group could significantly upregulate the cell proliferation and intracellular ROS content; CoCl2 could increase the protein expression of HIF-1α in a concentration-dependent manner. The results indicate that CoCl2 can induce hypoxia to a certain extent, facilitate intracellular oxidative stress, and promote the cell proliferation of primary PASMCs in a dose and time-dependent manner. Additionally, the proliferation effect of CoCl2 is weaker than gas hypoxia.
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