Background:Glucocorticoids(GCs)are widely used in acute and critical illnesses,but long-term and high-dose use of GCs can cause several vascular side effects.However,the underlying mechanisms are not well-understood.F...Background:Glucocorticoids(GCs)are widely used in acute and critical illnesses,but long-term and high-dose use of GCs can cause several vascular side effects.However,the underlying mechanisms are not well-understood.Ferroptosis,a novel form of reactive oxygen species(ROS)-dependent cell death,is characterized by intracellular iron accumulation and lipid peroxidation.NADPH oxidase 4(NOX4)is a major source of ROS.The roles of ferroptosis and NOX4 in GC-induced endothelial injury remain unknown.Methods:Human umbilical vein endothelial cells(HUVECs)were exposed to varying concentrations of dexamethasone(DEX)to evaluate ferroptosis and NOX4 expression.Further mechanistic studies were conducted using NOX4-overexpressing adenovirus(Ad-NOX4),NOX4 small interfering RNA(siRNA),ferrostatin-1(FER-1),and erastin.Results:Our findings demonstrate that DEX induces ferroptosis in HUVECs.Inhibition of ferroptosis with FER-1 prevents DEX-induced reduction in HUVEC viability.Furthermore,DEX treatment increases NOX4 expression in HUVECs,and NOX4 overexpression with AdNOX4 promotes ferroptosis.NOX4 knockdown with siRNA suppresses DEX-induced ROS production,lipid peroxidation,and ferroptosis,thereby improving the viability,angiogenesis,and migration capacity of DEX-treated HUVECs.However,the protective effect of NOX4 knockdown is negated by the reactivation of ferroptosis with erastin.Conclusion:GC-induced endothelial cell ferroptosis occurs through NOX4-mediated ROS production and lipid peroxidation,leading to cell death,impaired angiogenesis,and migration dysfunction.Inhibition of ferroptosis and NOX4 knockdown ameliorate GC-induced endothelial damage and dysfunction.展开更多
基金supported by the China Postdoctoral Science Foundation(2019M652399)National Natural Science Foundation of China(No.81600302)+2 种基金Shandong Natural Science Foundation(ZR2020QH336 and No.ZR2016HB01)Xinxin Green Valley Microcirculation Research Fund(XXLG07)the Chinese Ethnic Medicine Association(2023ZY086-46).
文摘Background:Glucocorticoids(GCs)are widely used in acute and critical illnesses,but long-term and high-dose use of GCs can cause several vascular side effects.However,the underlying mechanisms are not well-understood.Ferroptosis,a novel form of reactive oxygen species(ROS)-dependent cell death,is characterized by intracellular iron accumulation and lipid peroxidation.NADPH oxidase 4(NOX4)is a major source of ROS.The roles of ferroptosis and NOX4 in GC-induced endothelial injury remain unknown.Methods:Human umbilical vein endothelial cells(HUVECs)were exposed to varying concentrations of dexamethasone(DEX)to evaluate ferroptosis and NOX4 expression.Further mechanistic studies were conducted using NOX4-overexpressing adenovirus(Ad-NOX4),NOX4 small interfering RNA(siRNA),ferrostatin-1(FER-1),and erastin.Results:Our findings demonstrate that DEX induces ferroptosis in HUVECs.Inhibition of ferroptosis with FER-1 prevents DEX-induced reduction in HUVEC viability.Furthermore,DEX treatment increases NOX4 expression in HUVECs,and NOX4 overexpression with AdNOX4 promotes ferroptosis.NOX4 knockdown with siRNA suppresses DEX-induced ROS production,lipid peroxidation,and ferroptosis,thereby improving the viability,angiogenesis,and migration capacity of DEX-treated HUVECs.However,the protective effect of NOX4 knockdown is negated by the reactivation of ferroptosis with erastin.Conclusion:GC-induced endothelial cell ferroptosis occurs through NOX4-mediated ROS production and lipid peroxidation,leading to cell death,impaired angiogenesis,and migration dysfunction.Inhibition of ferroptosis and NOX4 knockdown ameliorate GC-induced endothelial damage and dysfunction.
