胰高血糖素样肽1受体激动剂对高糖高脂环境下大鼠心肌细胞损伤的改善作用及其铁死亡机制  

Improvement effect of glucagon-like peptide-1 receptor agonist on injury of cardiomyocytes in rats in hyperglycemia and hyperlipidemia environment and its iron death mechanism

作　　者：孟根托娅[1] 袁向珍[1] 解晓江[2] 成玲[3] 刘苗 MENGGENTUOYA;YUAN Xiangzhen;XIE Xiaojiang;CHENG Ling;LIU Miao(Physical Examination Center,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China;Department of Cardiovascular Medicine,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China;Medical Department,Affiliated Hospital,Inner Mongolia Medical University,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院体检中心,内蒙古呼和浩特010050 [2]内蒙古医科大学附属医院心血管内科,内蒙古呼和浩特010050 [3]内蒙古医科大学附属医院医务部,内蒙古呼和浩特010050

出　　处：《吉林大学学报（医学版）》2023年第4期823-831,共9页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金项目(81560045);内蒙古自治区科技厅自然科学基金项目(2020MS08145)。

摘　　要：目的:探讨胰高血糖素样肽1受体(GLP-1R)激动剂Exendin-4 (E4)对高糖高脂(HGHL)环境下心肌细胞损伤的影响,阐明其相关机制。方法:大鼠心肌H9C2细胞分为对照组、HGHL组、HGHL+E4组和HGHL+E4+铁死亡激活剂(Erastin)组,H9C2细胞采用33 mmol·L^(-1)葡萄糖和500μmol·L^(-1)棕榈酸脂诱导HGHL模型,HGHL+E4组和HGHL+E4+Erastin组H9C2细胞在诱导形成HGHL细胞模型后分别采用100 nmol·L^(-1) E4和5μmol·L^(-1) Erastin进行处理,CCK-8法检测各组细胞存活率,Hoechst 33258荧光染色法观察各组细胞凋亡情况,流式细胞术检测各组细胞凋亡率,2’,7’-二氢二氯荧光素二乙酸酯(DCFH-DA)荧光探针检测各组细胞中活性氧(ROS)水平,采用相关试剂盒检测各组细胞中谷胱甘肽(GSH)和丙二醛(MDA)水平,JC-1染色法检测各组细胞中线粒体膜电位(MMP)水平,FerroOrange荧光探针检测各组细胞中铁离子(Fe^(2+))水平,Westernblotting法检测各组细胞中谷胱甘肽过氧化物酶4 (GPX4)和溶质载体家族7成员11(SLC7A11)蛋白表达水平。结果:与对照组比较,HGHL组细胞存活率明显降低(P<0.05),细胞出现碎裂和皱缩,呈典型凋亡表现,细胞凋亡率及细胞中ROS和MDA水平明显升高(P<0.05),细胞中GSH和MMP水平明显降低(P<0.05),Fe^(2+)水平明显升高(P<0.05),GPX4和SLC7A11蛋白表达水平明显降低(P<0.05);与HGHL组比较,HGHL+E4组细胞存活率明显升高(P<0.05),凋亡细胞减少,细胞凋亡率及细胞中ROS和MDA水平明显降低(P<0.05),细胞中GSH和MMP水平明显升高(P<0.05),Fe^(2+)水平明显降低(P<0.05),GPX4和SLC7A11蛋白表达水平明显升高(P<0.05);与HGHL+E4组比较,HGHL+E4+Erastin组细胞存活率明显降低(P<0.05),细胞碎裂和皱缩现象明显减轻,细胞凋亡率及细胞中ROS和MDA水平明显升高(P<0.05),细胞中GSH和MMP水平明显降低(P<0.05),Fe^(2+)水平明显升高(P<0.05),GPX4和SLC7A11蛋白表达水平明显降低(P<0.05)。结论:GLP-1R激动剂能够减轻HGHL环境下心肌细�Objective:To disuss the effect of glucagon like peptide-1 receptor(GLP-1R)agonist Exendin-4(E4)on injury of cardiomyocytes in the hyperglycemia and hyperlipidemia(HGHL)environment,and to clarify its related mechanism.Methods:The H9C2 cardiomyocytes of the rats were divided into control group,HGHL group,HGHL+E4 group,HGHL+E4+iron death agonist(Erastin)group.The HGHL model of the H9C2 cells was induced by 33 mmo·l L^(-1) glucose and 500μmol·L^(-1) palmitate,and the cells in HGHL+E4 group and HGHL+E4+Erastin group were treated with 100 nmol·L^(-1) E4 and 5μmol·L^(-1) Erastin.CCK-8 assay was used to detect the survival rates of cells in various groups;Hoechst 33258 fluorescence staining was used to observe the apoptosis of cells in various groups;flow cytometry was used to detected the apoptotic rates of cells in various groups;2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)fluorescence probe was used to detect the levels of reactive oxygen species(ROS)in the cells in various groups;the corresponding kit was used to detect the levels of glutathione(GSH)and malondialdehyde(MDA)in cells in various groups;JC-1 staining was used to detect the mitochondrial membrane potential(MMP)in cells in various groups;FerroOrange fluorescence probe was used to detect the levels of Fe^(2+)in the cells in various groups;Western blotting method was used to detect the expression levels of glutathione peroxidase 4(GPX4)and solute carrier family 7 member 11(SLC7A11)in the cells in various groups.Results:Compared with control group,the survival rate of cells in HGHL group was decreased(P<0.05),and the cells were fragmented and collapsed,which showed apoptotic morphology;the apoptotic rate and the levels of ROS and MDA in the cells were increased(P<0.05),the levels of GSH and MMP were decreased(P<0.05),the level of Fe^(2+)was increased(P<0.05),and the expression levels of GPX4 and SLC7A11 proteins were decreased(P<0.05).Compared with HGHL group,the survival rate of the cells in HGHL+E4 group was increased(P<0.05),and the apoptosis

关 键 词：高糖高脂 心肌细胞 胰高血糖素样肽1受体激动剂 EXENDIN-4 铁死亡 

分 类 号：R587.2[医药卫生—内分泌]