Zhu Lijun, Feng Ziren, Zhang Jing, Du Lin, Meng Aiguo
Department of Laboratory Medicine, Affiliated hospital of North China university of science and technology, 063000, Tangshan, China.
Neurochem Res. 2023 May;48(5):1370-1381. doi: 10.1007/s11064-022-03826-3. Epub 2022 Dec 2.
Cerebral ischemia-reperfusion (I/R) injury is an inevitable issue in the treatment of ischemic stroke, which has a high disability rate and seriously threatens the living quality of patients. Previous studies have demonstrated that ferroptosis, which plays a crucial role in ischemia-reperfusion injury, can be accelerated by microRNA-27a (miR-27a). However, the mechanism by which miR-27a regulates ferroptosis in cerebral ischemia-reperfusion injury remains unknown. In this study, Male Sprague-Dawley rats were subjected to a middle cerebral artery occlusion (MCAO), then restored blood flow. Neurological function score and TTC staining were used to evaluate brain tissue injury and the infarct volume. The relative expression level of miR-27a was detected by qPCR. The relative expression levels of glutathione peroxidase 4(GPx4), solute carrier family 7 member 11 (SLC7A11) proteins were analyzed by Western Blot. The contents of GSH, Fe and malonaldehyde (MDA) were detected by corresponding detection kits, and the target gene of miR-27a was confirmed by dual luciferase reporter gene technique. It was found the relative expression level of miR-27a was increased and ferroptosis was aggravated as reperfusion time went by. Also, brain tissue injury and ferroptosis were exacerbated with agomiR-27a intervention, while these effects were reversed with antagomiR-27a intervention. In addition, the combined intervention of agomiR-27a and Fer-1 alleviated the brain tissue injury and ferroptosis. The results of dual luciferase reporter gene technique indicated SLC7A11 as the target gene of miR-27a. In the current study, miR-27a upregulates ferroptosis to aggravate cerebral ischemia-reperfusion injury by SLC7A11.
脑缺血再灌注(I/R)损伤是缺血性脑卒中治疗中不可避免的问题,其致残率高,严重威胁患者的生活质量。既往研究表明,在缺血再灌注损伤中起关键作用的铁死亡可被微小RNA-27a(miR-27a)加速。然而,miR-27a在脑缺血再灌注损伤中调节铁死亡的机制尚不清楚。在本研究中,雄性Sprague-Dawley大鼠接受大脑中动脉闭塞(MCAO),然后恢复血流。采用神经功能评分和TTC染色评估脑组织损伤及梗死体积。通过qPCR检测miR-27a的相对表达水平。采用蛋白质免疫印迹法分析谷胱甘肽过氧化物酶4(GPx4)、溶质载体家族7成员11(SLC7A11)蛋白的相对表达水平。用相应检测试剂盒检测谷胱甘肽(GSH)、铁和丙二醛(MDA)的含量,并通过双荧光素酶报告基因技术确定miR-27a的靶基因。结果发现,随着再灌注时间的延长,miR-27a的相对表达水平升高,铁死亡加重。此外,agomiR-27a干预可加重脑组织损伤和铁死亡,而antagomiR-27a干预可逆转这些效应。此外,agomiR-27a与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技术结果表明SLC7A11是miR-27a的靶基因。在本研究中,miR-27a通过SLC7A11上调铁死亡,加重脑缺血再灌注损伤。