H9c2 cells were incubated with a normal medium in a cell incubator for 24 h. Cells were then exposed to hypoxic conditions (oxygen deprivation, 1% O2) for 24 h in a culture medium with lower glucose and 1% FBS. After hypoxia, the cells were oxygenated under a normal oxygen concentration (reoxygenation) for 24 h in a normal medium. According to the previous study, propofol (Sigma-Aldrich, United States) was added respectively to the cells 1 h before and during the hypoxia-reoxygenation with different concentrations (5, 10, and 20 μM) (Zhao et al., 2015 (link)). U0126 (Beyotime, Haimen, China), MEK1/2 inhibitor, was added to the cells during OGD/R at a concentration of 20 μM. After H9c2 cells were incubated with a normal medium in cell incubator for 24 h, propofol and caspase inhibitor Z-VAD-FMK (Beyotime, Haimen, China) were respectively added to the cells for 24 h.
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