利拉鲁肽通过调节SIRT1改善高糖诱导的大鼠H9c2细胞氧化应激损伤

Liraglutide ameliorates high glucose-induced oxidative stress injury in rat H9c2 cells through modulation of SIRT1

目的探究利拉鲁肽(liraglutide, LRG)对高糖诱导的心肌细胞(H9c2)氧化应激损伤的影响及其潜在机制。方法采用高糖处理H9c2细胞24 h建立心肌细胞体外损伤模型, 给予不同浓度利拉鲁肽(10、100、1 000 nmol/L)干预, 用CCK-8检测细胞活力, 倒置显微镜观察细胞形态结构的改变。利拉鲁肽(100 nmol/L)干预高糖处理H9c2细胞24 h后检测细胞上清液中乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛含量;RT-PCR和Western印迹法检测沉默信息调节因子1(SIRT1)和叉头转录因子1(FOXO1)mRNA及蛋白水平;Western印迹法检测FOXO1蛋白乙酰化水平;应用小干扰RNA(siRNA)技术沉默SIRT1的H9c2细胞株验证SIRT1在其中的作用。结果与对照组相比, 高糖组细胞活力降低, 细胞结构受损, 细胞上清液中LDH、丙二醛含量显著升高, SOD含量降低, 氧化应激加重, SIRT1表达降低, FOXO1乙酰化水平升高(均P<0.05);与高糖组比较, 给予利拉鲁肽干预后细胞活力升高, 心肌细胞结构形态和氧化应激水平改善, SIRT1表达升高, FOXO1乙酰化水平降低(均P<0.05);当SIRT1下调后, 利拉鲁肽的上述保护效应减弱(均P<0.05)。结论利拉鲁肽对高糖诱导的H9c2细胞氧化应激损伤具有一定的保护作用, 其机制可能与上调SIRT1的表达水平有关。

Objective To investigate the effect of liraglutide(LRG)on high glucose-induced oxidative stress injury in(H9c2)cardiomyocytes and its underlying mechanisms.Methods A high glucose treatment was applied to H9c2 cells for 24 hours to establish an in vitro model of myocardial cell injury.Different concentrations of liraglutide(10,100,1000 nmol/L)were administered for intervention.Cell viability was evaluated using the CCK-8 assay,and changes in cell morphology were observed under an inverted microscope.After 24 hours of liraglutide(100 nmol/L)intervention following high glucose treatment,the levels of lactate dehydrogenase(LDH),superoxide dismutase(SOD),and malondialdehyde(MDA)in the cell supernatant were measured.RT-PCR and Western blotting were used to detect the mRNA and protein levels of silent information regulator factor 1(SIRT1)and forkhead box protein O1(FOXO1).Western blotting was also used to assess the acetylation level of FOXO1 protein.Small interfering RNA(siRNA)technology was employed to silence SIRT1 in H9c2 cells to confirm its role in the study.Results Compared to the control group,the high glucose group showed decreased cell viability,cell structure damage,increased levels of LDH and MDA in the cell supernatant,decreased SOD levels,aggravated oxidative stress,decreased SIRT1 expression,and increased acetylation level of FOXO1(all P<0.05).Compared to the high glucose group,liraglutide intervention resulted in increased cell viability,improved cardiac cell morphology,reduced oxidative stress levels,increased SIRT1 expression,and decreased acetylation level of FOXO1(all P<0.05).When SIRT1 was downregulated,the protective effects of liraglutide were weakened(all P<0.05).Conclusions Liraglutide has a protective effect against high glucose-induced oxidative stress injury in H9c2 cells,which may be associated with the upregulation of SIRT1 expression.