miR-29c-5p通过上调LRP6调控铁死亡减轻脑梗死缺血再灌注损伤

miR-29c-5p Aggravates Ischemic Stroke Reperfusion Injury by Promoting Ferroptosis via Downregulation of LRP6

目的:探究miR-29c-5p在脑缺血-再灌注损伤中调控铁死亡的机制。方法:在这项研究中,雄性SD大鼠接受大脑中动脉闭塞(MCAO)手术,随后恢复血液流动。采用神经功能评分和TTC染色来评估脑组织损伤和梗死体积。采用qPCR方法检测miR-29c-5p的相对表达水平。通过Western Blot检测谷胱甘肽过氧化物酶4(GPx4)、低密度脂蛋白受体相关蛋白6(LRP6)的表达水平。采用相应的检测试剂盒检测GSH、Fe和丙二醛(MDA)的含量,并通过双荧光素酶报告基因实验验证miR-29c-5p的靶基因。结果:随着大鼠缺血再灌注时间的增加,miR-29c-5p的相对表达水平升高,铁死亡加重。此外,agomiR-29c-5p干预也加重了脑组织损伤和铁死亡,而antagomiR-29c-5p干预则使这些影响得到逆转。此外,agomiR-29c-5p和Fer-1联合干预可减轻脑组织损伤和铁死亡。双荧光素酶报告基因实验结果表明,LRP6是miR-29c-5p的靶基因。结论:在本研究中,miR-29c-5p通过负调控LRP6促进铁死亡进而加重大鼠脑缺血-再灌注损伤。

Objective:To investigate the mechanism of miR-29c-5p in regulating ferroptosis in cerebral ischemia-reperfusion injury.Methods:In this study,male SD rats were subjected to middle cerebral artery occlusion(MCAO)surgery and then reperfused.Neurological function scores and TTC staining were used to assess brain injury and infarct volume.qPCR was used to detect the relative expression level of miR-29c-5p.Western blot was used to detect the expression levels of glutathione peroxidase 4(GPx4)and low-density lipoprotein receptor-related protein 6(LRP6).The contents of GSH,Fe,and malondialdehyde(MDA)were detected using the corresponding assay kits,and dual luciferase reporter gene assay was used to verify the target gene of miR-29c-5p.Results:With the increase of ischemia-reperfusion time in rats,the relative expression level of miR-29c-5p increased and ferroptosis was aggravated.In addition,agomiR-29c-5p intervention also aggravated brain injury and ferroptosis,while antagomiR-29c-5p intervention reversed these effects.Furthermore,combined intervention of agomiR-29c-5p and Fer-1 alleviated brain injury and ferroptosis.The results of dual luciferase reporter gene assay showed that LRP6 is a target gene of miR-29c-5p.Conclusion:In this study,miR-29c-5p promotes ferroptosis and aggravates cerebral ischemia-reperfusion injury in rats by downregulating LRP6.