瑞芬太尼对心肌缺血再灌注损伤及铁死亡过程的影响及机制研究

Investigation on the Effect and Mechanism of Remifentanil on Myocardial Ischemia-Reperfusion Injury and Ferroptosis

目的分析瑞芬太尼对心肌缺血再灌注损伤及铁死亡过程的影响及相关分子机制。方法选取人心肌细胞AC16进行氧糖剥夺处理及复氧处理复制心肌缺血再灌注(ischaemia/reperfusion,I/R)损伤模型,根据实验目的将细胞分为Control组、I/R组、I/R+瑞芬太尼组、I/R+瑞芬太尼+pcDNA组以及I/R+瑞芬太尼+ACSL 5组。EdU实验分析细胞增殖、流式细胞术分析细胞凋亡、Fe 2+比色法测定试剂盒分析Fe 2+水平、GSH检测试剂盒分析GSH水平、蛋白免疫印迹实验分析蛋白表达。结果瑞芬太尼缓解I/R处理对心肌细胞增殖的抑制作用和对细胞凋亡和铁死亡的促进作用;瑞芬太尼在I/R诱导的AC16细胞中下调ACSL4表达;瑞芬太尼通过调控ACSL4减弱I/R诱导的心肌细胞AC16凋亡和铁死亡进程。结论瑞芬太尼通过抑制ACSL4表达缓解I/R诱导的心肌细胞凋亡和铁死亡。

Objective To analyze the effect of remifentanil on myocardial ischemia-reperfusion(I/R)injury and ferroptosisl and the related molecular mechanisms.Methods All experiments were conducted in human cardiomyocytes AC16.The cells were treated with oxygen and sugar deprivation followed by reoxygenation to simulate myocardial I/R injury.Cells were divided into Control group,I/R group,I/R+Remifentanil group,I/R+Remifentanil+pcDNA group,and I/R+Remifentanil+ACSL4 group according to the experimental purposes.EdU experiment was performed to analyze cell proliferation,flow cytometry was used to analyze cell apoptosis,Fe 2+colorimetric assay kit was conducted to determine the level of Fe 2+,GSH detection kit was used to analyze GSH level,while Western blot experiment was utilized to analyze protein expression.Results Remifentanil alleviated the inhibitory effect of I/R treatment on cardiomyocyte proliferation and the promotion of apoptosis and ferroptosisl.Remifentanil downregulates ACSL4 expression in I/R-induced AC16 cells.Remifentanil attenuated the process of I/R-induced apoptosis and ferroptosisl in AC16 cardiomyocytes by modulating ACSL4.Conclusion Remifentanil inhibited ACSL4 expression to alleviate I/R-induced apoptosis and ferroptosis of human cardiomyocytes.