抑制细胞迁移诱导蛋白对缺氧复氧诱导的微循环内皮细胞铁死亡的影响  

Impact of inhibiting cell migration-inducing protein on ferroptosis in microvascular endothelial cell induced by hypoxia-reoxygenation

作　　者：刘双[1] 刘根 陈静[1] 张博方[1] LIU Shuang;LIU Gen;CHEN Jing;ZHANG Bofang(Department of Cardiology,Renmin Hospital of Wuhan University Cardiovascular Research Institute Wuhan University Hubei Key Laboratory of Cardiology,Hubei Province,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院心内科心血管病湖北省重点实验室武汉大学心血管病研究所,湖北武汉430060

出　　处：《中国医药导报》2024年第28期1-6,共6页China Medical Herald

基　　金：国家自然科学基金资助项目(82100287);湖北省自然科学基金青年项目(2021CFB110)。

摘　　要：目的探讨抑制细胞迁移诱导蛋白(CEMIP)对缺氧复氧诱导的心脏微循环内皮细胞(CMEC)铁死亡的作用及其机制。方法分离培养原代大鼠CMEC,并分为正常组和缺氧复氧组(H/R组)。根据是否应用腺病毒下调CEMIP表达,进一步分为转染对照病毒+缺氧复氧组(sh-con+H/R组)和转染CEMIP下调病毒+缺氧复氧组(sh CEMIP+H/R组)。运用RNA测序技术筛选正常组与H/R组差异基因。采用CCK8法和Transwell法检测细胞的增殖和迁移能力,铁含量测定试剂盒检测铁离子含量,Hoechst 33342/PI双荧光染色检测CMEC的死亡,采用q PCR检测PTGS2 m RNA,免疫蛋白印记法检测CEMIP和SLC7A11蛋白的水平。结果RNA测序分析显示H/R组中CEMIP升高显著。与正常组比较,H/R组CMEC中CEMIP升高,增殖和迁移能力降低,同时细胞死亡增加,活性铁的含量、PTGS2 m RNA升高、SLC7A11蛋白降低(P<0.05)。此外,sh CEMIP+H/R组CMEC增殖、迁移能力和SLC7A11蛋白水平低于sh-con+H/R组,而活性铁含量、PTGS2水平高于sh-con+H/R组(P<0.05)。结论在缺氧复氧过程中,CEMIP可能靶向调控SLC7A11的表达来缓解CMEC的铁死亡。Objective To investigate the impact and mechanism of cell migration-inducing protein(CEMIP)on ferroptosis in cardiac microvascular endothelial cell(CMEC)induced by hypoxia-reoxygenation.Methods Primary rat CMEC were isolated and cultured,and they were divided into control group and hypoxia-reoxygenation group(H/R group).CEMIP expression was further divided into transfection control virus+hypoxia reoxygenation group(sh-con+H/R group)and transfection CEMIP down-regulated virus+hypoxia reoxygenation group(shCEMIP+H/R group)according to whether adenovirus was used to down-regulate CEMIP expression.RNA sequencing was employed to screen for differentially expressed genes between the control group and H/R group.Cell proliferation and migration were assessed using CCK8 and Transwell assays.Iron ion content was measured by iron assay kit.CMEC death was detected using Hoechst 33342/PI fluorescence staining.The levels of PTGS2 mRNA were determined by qPCR,and the expression of CEMIP and SLC7A11 proteins was analyzed by Western blot.Results RNA sequencing heatmaps and volcano plots showed a significant increase in CEMIP in the H/R group.Compared with normal group,CEMIP in CMEC in H/R group was increased,proliferation and migration ability was decreased,cell death was increased,active iron content,PTGS2 mRNA were increased,and SLC7A11 protein was decreased(P<0.05).In addition,the proliferation and migration capacity of CMEC and the protein level of SLC7A11 in shCEMIP+H/R group were lower than those in sh-con+H/R group,while the active iron content and PTGS2 level in SHCEMIP+H/R group were higher than those in sh-con+H/R group(P<0.05).Conclusion CEMIP may regulate SLC7A11 expression under hypoxic/reoxygenation conditions,thereby alleviating the ferroptosis of CMEC.

关 键 词：细胞迁移诱导蛋白 心脏微循环内皮细胞 缺氧复氧 铁死亡 

分 类 号：R54[医药卫生—心血管疾病]