中国病理生理杂志2025，Vol.41Issue(6)：1162-1169,8.DOI:10.3969/j.issn.1000-4718.2025.06.013
miR-351-5p在脂多糖诱导的心肌细胞铁死亡过程中的调控作用
Regulatory role of miR-351-5p in lipopolysaccharide-induced ferroptosis of cardiomyocytes
摘要
Abstract
AIM:This study aims to investigate the role of ferroptosis in the myocardium of mice with lipopoly-saccharide(LPS)-induced sepsis and in the injury of H9c2 rat cardiomyocytes,and to explore the regulatory function of microRNA-351-5p(miR-351-5p)in this context.METHODS:An in vivo model of sepsis-induced cardiomyopathy was established in mice through intraperitoneal injection of LPS.Twenty-four mice were randomly divided into negative control(NC)group,LPS group,and LPS+ferroptosis inhibitor ferrostatin-1(Fer-1)group.Hematoxylin-eosin(HE)staining was conducted to assess cardiac injury,and plasma levels of creatine kinase(CK)and lactate dehydrogenase(LDH)were also measured.Additionally,the levels of Fe2+and malondialdehyde(MDA)in plasma were quantified,and the mRNA levels of acyl-CoA synthetase long chain family member 4(ACSL4)and prostaglandin-endperoxide synthase 2(PTGS2)were de-tected by RT-qPCR.In vitro,H9c2 cardiomyocytes were stimulated with LPS to create cellular models,followed by treat-ment with Fer-1,inhibitor NC,or miR-351-5p inhibitor.Cell viability was evaluated using CCK8 assay,intracellular re-active oxygen species(ROS)were measured by flow cytometry,intracellular Fe2+levels were assessed using a fluorescence probe,and the protein expression of glutathione peroxidase 4(GPX4)and ACSL4 was analyzed by Western blot.The MDA and reduced glutathione(GSH)levels were measured using commercial kits.MicroRNA(miRNA)sequencing was performed on the LPS-stimulated H9c2 cardiomyocyte models,with differential miRNAs identified and subsequently vali-dated using RT-qPCR.RESULTS:The mice in LPS group exhibited significant myocardial tissue dysregulation com-pared with NC group,with enlarged space,increased plasma CK and LDH levels(P<0.05),elevated Fe2+and MDA levels in myocardial tissues(P<0.05),and increased mRNA levels of ACSL4 and PTGS2(P<0.05).In contrast,the mice in LPS+Fer-1 group demonstrated improved myocardial tissue structure,reduced space,decreased plasma CK and LDH levels(P<0.05),and lower Fe2+and MDA levels in myocardial tissues(P<0.05),along with decreased mRNA level of PTGS2(P<0.05).In H9c2 cardiomyocytes,cell viability,intracellular GSH level,and GPX4 protein level were significantly reduced in LPS group compared with NC group(P<0.05),while ROS,MDA,Fe2+,and ACSL4 protein levels were elevated(P<0.05).The cells in LPS+Fer-1 group showed increased viability,intracellular GSH level,and GPX4 protein level compared with LPS group(P<0.05),alongside reduced ROS,MDA,Fe2+,and ACSL4 levels(P<0.05).miRNA sequencing revealed a significant decrease in several miRNAs,with miR-351-5p showing the most pro-nounced reduction.In LPS+miR-351 inhibitor group,H9c2 cell viability significantly declined(P<0.05),and the levels of GSH and GPX4 were notably lowered(P<0.05),while ROS,MDA,Fe2+and ACSL4 protein levels were significantly elevated(P<0.05).However,in LPS+miR-351 inhibitor+Fer-1 group,the cell viability increased(P<0.05),and the GSH level rose significantly(P<0.05),with corresponding decreases in intracellular ROS,Fe2+and ACSL4 protein levels(P<0.05).CONCLUSION:Inhibition of ferroptosis attenuated sepsis-induced myocardial injury,and inhibition of miR-351-5p promotes sepsis-induced ferroptosis of H9c2 cardiomyocytes.关键词
脓毒症心肌病/铁死亡/微小RNA-351-5pKey words
sepsis-induced cardiomyopathy/ferroptosis/microRNA-351-5p分类
临床医学引用本文复制引用
张梦茹,彭燕芬,李晴雯,付丽珊,冉庆森,李丹丹,李保林..miR-351-5p在脂多糖诱导的心肌细胞铁死亡过程中的调控作用[J].中国病理生理杂志,2025,41(6):1162-1169,8.基金项目
深圳市坪山区卫生健康局及科技创新局基金资助项目(No.202115) （No.202115）
