山西医科大学学报2025，Vol.56Issue(9)：1016-1024,9.DOI:10.13753/j.issn.1007-6611.2025.09.005
Caveolin-1在氧糖剥夺诱导的人脑微血管内皮细胞铁死亡中的作用
Role of Caveolin-1 in ferroptosis of human brain microvascular endothelial cells induced by oxygen-glucose deprivation
摘要
Abstract
Objective To investigate whether ischemic injury in human brain microvascular endothelial cells is regulated by ferropto-sis and Caveolin-1(CAV1).Methods To investigate the effect of oxygen-glucose deprivation(OGD)on ferroptosis in human cerebral microvascular endothelial cells,the cells were divided into control group and OGD group.Cells in control group were cultured under normal conditions,while the cells in OGD group were cultured with a sugar-free medium under hypoxia for 6 h,and then reoxygena-tion with a sugar-containing medium for 12 h.FerroOrange fluorescent probe was employed to measure ferrous ion(Fe2+)level,and BODIPY(581/591)C11 fluorescent probe was used to quantify lipid peroxide content.To elucidate the role of ferroptosis in OGD,the cells were categorized into control group,OGD group,and OGD+deferoxamine group.After normal cells had adhered to the culture surface,the cells in OGD+deferoxamine group were treated with 1 μmol/L ferroptosis inhibitor deferoxamine,and then oxygen-glucose deprivation.The colony-forming ability of cells was assessed using colony formation assay.To explore the influence of CAV1 on ferrop-tosis during OGD in human cerebral microvascular endothelial cells,the cells were divided into control group,OGD group,OGD+empty vector group,and OGD+CAV1 overexpression group.The cells in OGD+empty vector group and OGD+CAV1 overexpression group were infected with empty lentivirus and CAV1 overexpression lentivirus before establishing the OGD model,respectively.The ef-ficacy of CAV1 overexpression lentivirus infection was evaluated by Western blotting.The colony-forming ability of cells was deter-mined using colony formation assay.The Fe2+level was measured with the FerroOrange fluorescent probe,and the lipid peroxide con-tent was quantified using the BODIPY(581/591)C11 fluorescent probe.The protein levels of glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and transferrin receptor(TFR)were detected by immunofluorescence.The glutathione(GSH)content was measured using a GSH/GSSG assay kit,and the reactive oxygen species(ROS)content was detected with the DCFH-DA fluorescent probe.Results Compared with control group,the cells in OGD group exhibited significantly elevated levels of Fe2+and lipid peroxides(P＜0.05),along with a marked reduction in CAV1 expression(P＜0.05).Additionally,the colony-forming ability of cells in OGD group was notably weaker than that in control group(P＜0.05).The cells in OGD+deferoxamine group demonstrated a sig-nificantly enhanced colony-forming ability compared to OGD group(P＜0.05).Compared with control group,the cells in OGD group showed significantly increased contents of Fe2+,lipid peroxides,ROS,and TFR protein(P＜0.05).Conversely,the cells in OGD+CAV1 overexpression group exhibited significantly decreased levels of Fe2+,lipid peroxides,ROS,and TFR protein compared to OGD group(P＜0.05).Moreover,the cells in OGD group exhibited significantly increased contents of GSH,GPX4 protein,and SLC7A11 protein compared to control group(P＜0.05).In contrast,the cells in OGD+CAV1 overexpression group showed significantly elevated levels of GSH,GPX4 protein,and SLC7A11 protein compared to OGD group(P＜0.05).Conclusion OGD can induce the ferropto-sis in human brain microvascular endothelial cells and suppress CAV 1 expression.Overexpression of CAV1 can inhibit OGD-induced ferroptosis in these cells.These findings suggest that the ischemic injury in human brain microvascular endothelial cells is regulated by ferroptosis and CAV1.关键词
人脑微血管内皮细胞/缺氧复氧/小窝蛋白1/氧糖剥夺/铁死亡/GPX4/SLC7A11Key words
human brain microvascular endothelial cells/hypoxia-reoxygenation/Caveolin-1/oxygen-glucose deprivation/ferroptosis/GPX4/SLC7A11分类
基础医学引用本文复制引用
邹荣基,喻芳芳,刘腾飞,李丹霞,张俊斌,贾卓鹏..Caveolin-1在氧糖剥夺诱导的人脑微血管内皮细胞铁死亡中的作用[J].山西医科大学学报,2025,56(9):1016-1024,9.基金项目
西藏自治区自然科学基金组团式援藏医学项目(XZ2024ZR-ZY121(Z)) （XZ2024ZR-ZY121(Z）
