罗格列酮调控沉默信息调节因子1对高糖处理的心肌微血管内皮细胞增殖和迁移的影响

Effects of rosiglitazoneon on the proliferation and migration of cardiac microvascular endothelial cells treated with high glucose by regulating Sirt1

目的:探讨罗格列酮对高糖处理的心肌微血管内皮细胞(CMEC)增殖、迁移的影响和分子机制。方法:将大鼠CMEC分为对照组(葡萄糖5.5 mmol/L)、高糖对照组(葡萄糖33 mmol/L)、实验1组(高糖+罗格列酮5μmol/L)、实验2组(高糖+罗格列酮10μmol/L)、实验3组(高糖+罗格列酮20μmol/L)。细胞计数试剂盒8(CCK-8)法检测细胞增殖活力;Transwell实验检测细胞迁移能力;Western blot检测P21、上皮细胞钙粘蛋白(E-cadherin)、沉默信息调节因子1(Sirt1)的蛋白表达水平;实时荧光定量聚合酶链反应检测Sirt1的mRNA表达水平。将Sirt1小干扰RNA(si-Sirt1)转染CMEC,经高糖和罗格列酮20μmol/L处理后,采用上述方法检测细胞增殖和迁移能力。结果:与对照组比较,高糖对照组CMEC增殖和迁移能力降低,P21和E-cadherin表达增加,Sirt1表达降低(P均<0.05)。与高糖对照组比较,高糖联合罗格列酮处理后CMEC增殖和迁移能力增加,P21和E-cadherin表达降低,Sirt1表达增加(P均<0.05)。与高糖联合罗格列酮处理比较,转染si-Sirt1并经高糖联合罗格列酮处理后CMEC增殖和迁移能力降低,P21和E-cadherin表达增加(P均<0.05)。结论:罗格列酮通过上调Sirt1表达促进高糖处理的CMEC增殖和迁移。

Objective:To investigate the effects and molecular mechanism of rosiglitazone on proliferation and migration cardiac microvascular endothelial cells(CMECs)treated by high glucose.Methods:Rat CMECs were divided into control group(5.5 mmol/L glucose),high glucose control group(33 mmol/L glucose),experimental group 1(33 mmol/L glucose+5μmol/L rosiglitazone),experimental group 2(33 mmol/L glucose+10μmol/L rosiglitazone)and experiment 3 group(33 mmol/L glucose+20μmol/L rosiglitazone).Cell counting kit 8(CCK-8)was used to detect cell proliferation activity.Transwell test was used to detect cell migration ability.Western blot was used to detect the expression levels of P21,epithelial cadherin(E-cadherin)and silent information transcriptional regulator 1(Sirt1).Real-time fluorescent quantitative PCR was used to detect Sirt1 mRNA expression.The Sirt1 small interfering RNA(si-Sirt1)was transfected into CMECs,which was exposed to high glucose and 20μmol/L rosiglitazone,and then the cell proliferation and migration abilities were measured by the above method.Results:Compared with the control group,the proliferation and migration abilities of CMECs and the expression level of Sirt1 were decreased in high glucose control group,while the expression levels of P21 and E-cadherin were increased(all P<0.05).Compared with the high glucose control group,the proliferation and migration abilities of CMECs and the expression levels of Sirt1 were increased after treated with high glucose with rosiglitazone,while the expression levels of P21 and E-cadherin were decreased(all P<0.05).Compared with high glucose combined with rosiglitazone treatment group,the proliferation and migration abilities of CMECs were declined,and the expression levels of P21 and E-cadherin were increased after transfection of si-Sirt1(all P<0.05).Conclusions:Rosiglitazone promotes the proliferation and migration abilities of CMECs treated with high glucose by up-regulating Sirt1 expression.