Abstract:

BACKGROUND: Studies have shown that the apoptosis of human coronary artery endothelial cells is involved in the generation and development of atherosclerosis.
OBJECTIVE: To investigate the inductive effect of tumor necrosis factor alpha on the damage of human coronary artery endothelial cells.
METHODS: First, human coronary artery endothelial cells c-12221 in the logarithmic growth phase were selected. Next, the cells were added into 0 (control), 200, 400 and 600 mg/L tumor necrosis factor alpha culture medium. Then, the changes of cell proliferation rate, morphological changes of apoptosis cells, changes of cell apoptosis rate and mitochondrial membrane potential were detected by MTT assay, Hoechst 33258/propidium iodide staining, fluorescence microscopy, Annexin V-FITC and PI-double staining flow cytometry assay and high content live cell imaging system, respectively.
RESULTS AND CONCLUSION: MTT results showed that tumor necrosis factor alpha had potential inhibitory effects on human coronary artery endothelial cells proliferation in a dose-dependent manner. Hoechst 33258/propidium iodide staining indicated that characteristic change of apoptotic cells could be seen clearly, such as chromatin condensation, nuclear fragmentation into pieces. Besides, cell apoptosis rate of tumor necrosis factor alpha with different concentrations was higher than that of 0 mg/L tumor necrosis factor alpha (P < 0.05); their mitochondrial membrane potential was lower than that of the control and in a dose-dependent manner (P < 0.05). These results suggest that tumor necrosis factor alpha can promote the apoptosis of human coronary artery endothelial cells in a dose- and time-dependent manner and inhibit human coronary artery endothelial cell proliferation. The underlying mechanisms are related to mitochondrial apoptosis pathway.