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
作者
孟根托娅
袁向珍
解晓江
成玲
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)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
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)。
