Pulmonary microvascular endothelial cells (PMVECs; Sunncell, Wuhan, China) were cultured in 89% DMEM (Sunncell) supplemented with 10% FBS (Sigma‐Aldrich), 0.5% penicillin and 0.5% streptomycin in a humidified atmosphere with 5% CO2 at 37 °C.
Cells were grouped into the control, the oxygen/glucose deprivation and reoxygenation (OGD/R), the OGD/R + emodin and the OGD/R + Bay‐117082. The PMVECs were subjected to PBS washing for three times, fed medium without glucose and serum, and then placed for 1 h at 37 °C in a Whitley H35 Hypoxystation (Hua Yue, Guangzhou, China) in an atmosphere of 1% O2, 5% CO2, and 94% N2. Subsequently, the cells were cultured for 4 h at 37 °C in glucose‐containing medium in an atmosphere of 5% CO2 and 95% O2.30 The cells in the OGD/R + emodin group were pretreated with 40‐μM emodin 1 h before reoxygenation. In the OGD/R + Bay‐117082 group, the cells were pretreated with 5‐μM Bay‐117082 (Selleck Chemicals, Shanghai, China) for 1 h before reoxygenation.
Cells were grouped into the control, the oxygen/glucose deprivation and reoxygenation (OGD/R), the OGD/R + emodin and the OGD/R + Bay‐117082. The PMVECs were subjected to PBS washing for three times, fed medium without glucose and serum, and then placed for 1 h at 37 °C in a Whitley H35 Hypoxystation (Hua Yue, Guangzhou, China) in an atmosphere of 1% O2, 5% CO2, and 94% N2. Subsequently, the cells were cultured for 4 h at 37 °C in glucose‐containing medium in an atmosphere of 5% CO2 and 95% O2.
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