铁死亡参与二氧化硅诱导血管内皮细胞损伤的作用研究  

Study on the Role of Ferroptosis in SiO2 Induced Injury of Vascular Endothelial Cells

作　　者：王秋实 孙静 孙岳 畅思容 王凯[1] 张娜[1] 彭泽钦 刘志宏[1] WANG Qiushi;SUN Jing;SUN Yue;CHANG Sirong;WANG Kai;ZHANG Na;PENG Zeqin;LIU Zhihong(School of Public Health,Ningxia Medical University,Yinchuan 750004,China;Ning An Hospital of Ningxia Hui Autonomous Region,Yinchuan 750011,China;Chongqing Medical University,Chongqing 404100,China)

机构地区：[1]宁夏医科大学公共卫生学院,银川750004 [2]宁夏回族自治区宁安医院,银川750011 [3]重庆医科大学,重庆404100

出　　处：《宁夏医科大学学报》2024年第12期1189-1196,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(82360639;82060264;82260142);宁夏回族自治区重点研发计划项目(2021BEB04059);宁夏自然科学基金项目(2022AAC03141,2024AAC03241)。

摘　　要：目的 阐明铁死亡参与二氧化硅(SiO_(2))诱导的血管内皮细胞损伤过程,为矽肺防治提供依据。方法 将人脐静脉内皮细胞(HUVECs)作为实验对象,分为对照(Control)组、SiO_(2)组、SiO_(2)+铁死亡抑制剂(Fer-1)组。q RT-PCR及Western blot评价铁死亡及氧化应激相关因子m RNA、蛋白表达水平;使用荧光探针通过免疫荧光观察分析活性氧(ROS)表达水平与细胞损伤水平。结果 与Control组比较,随着SiO_(2)浓度升高,HUVECs细胞活力降低(P<0.05),且Si O_(2)组细胞损伤水平升高(P<0.05);与Control组比较,Si O_(2)组HUVECs中铁离子和丙二醛(MDA)含量均增加(P均<0.05),ROS增加(P<0.05),GPX4和Ferritin的蛋白及m RNA相对表达水平均降低(P均<0.05),白细胞介素(IL)-1α、IL-6、肿瘤坏死因子(TNF)-α的m RNA相对表达水平均上调(P均<0.05);与SiO_(2)组比较,SiO_(2)+Fer-1组HUVECs中铁离子和MDA含量均降低(P均<0.05),GPX4和Ferritin的蛋白及m RNA相对表达水平均上升(P均<0.05),IL-1α、IL-6、TNF-α的m RNA相对表达水平均下调(P均<0.05),ROS下降(P<0.05),细胞损伤水平降低(P<0.05)。结论 铁死亡参与了SiO_(2)诱导的血管内皮细胞损伤及炎性反应过程,SiO_(2)导致的血管内皮细胞损伤及炎性反应能被Fer-1缓解。Objective To clarify the involvement of ferroptosis in the process of vascular endothelial cell damage induced by silica(SiO_(2))and provide a basis for the prevention and treatment of silicosis.Methods Human umbilical vein endothelial cells(HUVECs)were used as experimental subjects and divided into Control group,SiO_(2) group,SiO_(2)+ferrostatin-1(Fer-1)group.RT-qPCR and Western blot were employed to evaluate the mRNA and protein expression levels of factors related to ferroptosis and oxidative stress.Additionally,fluorescent probes were utilized to analyze the expression levels of reactive oxygen species(ROS)and cellular damage through immunofluorescence observation.Results Compared with the Control group,with the increase of SiO_(2) concentration,the cell viability of HUVECs decreased(P<0.05),and the cell damage level in the SiO_(2) group increased(P<0.05).Compared with the Control group,the iron ion and malondialdehyde(MDA)contents in HUVECs in the SiO_(2) group increased(P all<0.05),ROS increased(P<0.05),the protein and mRNA relative expression levels of GPX4 and Ferritin decreased(P all<0.05),and the expression level of interleukin(IL)-1α,IL-6 and tumor necrosis factor(TNF)-αwas upregulated(P all<0.05).Compared with the SiO_(2) group,the contents of iron ions and MDA in HUVECs in the SiO_(2)+Fer-1 group were decreased(P<0.05),the relative expression levels of GPX4 and Ferritin protein and mRNA were increased(P all<0.05),the expression levels of IL-1α,IL-6,and TNF-αwere downregulated(P all<0.05),ROS decreased(P<0.05),and the level of cell damage was reduced(P<0.05).Conclusion Ferroptosis is implicated in the mechanistic pathway of SiO_(2)-induced injury and inflammatory response within HUVECs.Specifically,the detrimental effects and inflammatory cascade triggered in HUVECs as a consequence of silica exposure can be notably ameliorated through the application of the Fer-1.

关 键 词：矽肺 铁死亡 血管内皮细胞 脂质过氧化 铁死亡抑制剂 

分 类 号：R363[医药卫生—病理学]