格列本脲对H_(2)O_(2)诱导的缺氧心肌细胞水肿及SUR1-TRPM4通道和AQP4表达的影响  

Effects of glibenclamide on H_(2)O_(2)-induced edema of hypoxic cardiomyocytes,the SUR1-TRPM4 channel and expression level of AQP4

作　　者：沈莹[1] 曾永孝 邓蓉 谭倩 雷超[1] SHEN Ying;ZENG Yongxiao;DENG Rong(Department of Geriatrics,Liyuan Hospital of Tongji Medical College of Huazhong University of Science&Technology,Hubei,Wuhan 430077,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院老年病科,武汉市430077 [2]华中科技大学同济医学院附属梨园医院内分泌科,武汉市430077 [3]湖北省荆门市中医医院(石化医院)神经内科

出　　处：《河北医药》2024年第24期3702-3706,共5页Hebei Medical Journal

基　　金：湖北省卫生健康立项项目(编号:WJ2023F010)。

摘　　要：目的探讨格列本脲对H_(2)O_(2)诱导的缺氧心肌细胞水肿及SUR1-TRPM4通道和AQP4表达的影响。方法将大鼠心肌H9C2细胞分为对照组、格列本脲低剂量组、格列本脲高剂量组、格列本脲高剂量+CIM2016(SUR1-TRPM4通道激活剂)组;MTT染色法检测细胞存活率;透射电镜观察细胞水肿超微结构变化;试剂盒检测MDA、SOD、CAT含量;qRT-PCR检测5组细胞SUR1、TRPM4、AQP4的基因表达水平;western blot检测细胞中SUR1、TRPM4、AQP4、BAX、BCL-2蛋白表达水平。结果与对照组比较,模型组H9C2细胞体积明显肿胀增大,细胞存活率、细胞SOD、CAT活性、BCL-2蛋白表达显著降低(P<0.05),MDA含量、细胞SUR1 mRNA、TRPM4 mRNA、AQP4 mRNA表达,细胞SUR1、TRPM4、AQP4、BAX蛋白表达显著升高(P<0.05);与模型组比较,格列本脲低、高剂量组H9C2细胞体积肿胀程度减轻,细胞存活率、细胞SOD、CAT活性、BCL-2蛋白表达显著升高(P<0.05),MDA含量、细胞SUR1 mRNA、TRPM4 mRNA、AQP4 mRNA表达,细胞SUR1、TRPM4、AQP4、BAX蛋白表达显著降低(P<0.05);CIM2016可部分逆转格列本脲对H_(2)O_(2)诱导的缺氧心肌细胞水肿的改善作用(P<0.05)。结论格列本脲可改善H_(2)O_(2)诱导的缺氧心肌细胞水肿,提高细胞存活率,抑制SUR1-TRPM4通道和AQP4表达。Objective To investigate the effects of glibenclamide on H_(2)O_(2)-induced edema of hypoxic cardiomyocytes,the sulfonylurea receptor 1-transient receptor potential melastatin 4(Sur1-Trpm4)channel and expression level of aquaporin-4(AQP4).Methods The rat cardiac cell line H9C2 was induced with blank control,H_(2)O_(2)+low-dose glibenclamide,H_(2)O_(2)+high-dose glibenclamide,and H_(2)O_(2)+high-dose glibenclamide+CIM2016(SUR1-TRPM4 channel activator).Cell survival and ultrastructural changes in cell edema were detected by MTT assay and transmission electron microscopy(TEM).Contents of malondialdehyde(MDA),superoxide dismutase(SOD),and catalase(CAT)were measured by commercial kits.Quantitative real-time polymerase chain reaction(QRT-PCR)was applied to detect the mRNA levels of SUR1,TRPM4,and AQP4.Western blot was applied to detect the protein levels of SUR1,TRPM4,AQP4,BAX,and BCL-2 proteins in cells.Results Compared with those of the control group,H_(2)O_(2)-induced H9C2 cells were greatly swollen in volume,with significantly lower survival rate,SOD and CAT activities,and protein level of Bcl-2,but higher MDA content,mRNA Levels of SUR1,TRPM4 and AQP4,and protein levels of SUR1,TRPM4,AQP4 and Bax(P<0.05).Compared with those induced with H_(2)O_(2),H9C2 cells induced with H_(2)O_(2)+low-dose/high-dose glibenclamide had alleviated volume swelling,significantly higher survival rate,SOD and CAT activities,and protein level of Bcl-2,but lower MDA content,mRNA Levels of SUR1,TRPM4 and AQP4,and protein levels of SUR1,TRPM4,AQP4 and Bax(P<0.05).CIM2016 partially reversed the role of glibenclamide in alleviating H_(2)O_(2)-induced swelling of H9C2 cells(P<0.05).Conclusion Glibenclamide can alleviate H_(2)O_(2)-induced edema of hypoxic cardiomyocytes,increase cell survival rate,and inhibit the SUR1-TRPM4 channel and expression level of AQP4.

关 键 词：格列本脲 H2O2 缺氧心肌细胞水肿 SUR1-TRPM4通道 AQP4 

分 类 号：R541[医药卫生—心血管疾病]