miRNA-137靶向SLC1A5调控铁死亡对大鼠脑缺血再灌注损伤的保护机制  

The protective mechanism of miRNA-137 targeting SLC1A5 regulates ferroptosis against cerebral ischemia-reperfusion injury in rats

作　　者：向军军[1,2] 赖菁菁[1] 莫雪妮[2] 陈炜[1] 胡跃强[1] 姚春[2] XIANG Jun-Jun;LAI Jing-Jing;MO Xue-Ni(The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,Guangxi,China;不详)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530023 [2]广西中医药大学

出　　处：《中国老年学杂志》2024年第24期5981-5986,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金(81973768);广西中医药大学2021年研究生教育创新计划项目(YCBXJ2021008)。

摘　　要：目的观察miRNA-137靶向溶质载体家族1成员(SLC1A)5对脑缺血再灌注损伤(CIRI)大鼠的神经功能影响及其调节铁死亡的可能机制。方法60只成年健康SD大鼠,随机分为6组:假手术(Sham)组、模型(Model)组、miR-137模拟剂(miR-137 mimic)组、miR-137抑制剂(miR-137 inhibitor)组,miR-137空载体病毒(miR-137 vector)组、Ferrostatin-1铁死亡抑制剂(Fer-1)组(5 mg/kg),于造模前1 w,各病毒组从侧脑室注入2μl病毒载体,连续7 d;在造模后第1、3、7天时采用Longa法评估大鼠神经功能,并在第7天神经功能评分后取材,生化试剂盒检测还原型谷胱甘肽(GSH)及二价铁离子(Fe^(2+))表达,qRT-聚合酶链反应(PCR)检测各组脑组织中miR-137、SLC1A5、溶质载体家族7成员(SLC7A)11及谷胱甘肽过氧化物酶(GPX)4表达水平,Western印迹检测SLC1A5、SLC7A11及GPX4蛋白表达,并采用电镜观察各组脑组织中神经元线粒体铁死亡的变化。结果与Sham组比较,Model组神经功能缺损评分、Fe^(2+)含量、SLC1A5 mRNA及蛋白表达明显升高,GSH含量、SLC7A11、GPX4 mRNA及蛋白表达明显下降(P<0.05,P<0.01),电镜下可见线粒体缩小、线粒体膜增厚、线粒体嵴溶解断裂;与Model组比较,miR-137 mimic组神经功能缺损评分、Fe^(2+)含量、SLC1A5 mRNA及蛋白表达显著下降,GSH含量、SLC7A11、GPX4 mRNA及蛋白表达显著升高(P<0.05,P<0.01),线粒体形态学损伤明显改善,表现为线粒体膜完整、线粒体嵴数目多、结构清晰;与miR-137 mimic组比较,miR-137 inhibitor与miR-137 vector组神经功能缺损评分、Fe^(2+)含量、SLC1A5 mRNA及蛋白表达明显升高,GSH含量、SLC7A11、GPX4 mRNA及蛋白表达明显下降(P<0.05,P<0.01),电镜下可见线粒体缩小、线粒体膜增厚、线粒体嵴溶解断裂;而miR-137 mimic组与Fer-1组各指标差异无统计学意义(P>0.05)。结论过表达miR-137对CIRI大鼠神经损伤具有保护作用,其机制可能与靶向抑制SLC1A5蛋白表达,上调抗氧化因子SLC7A11、GObjective To observe the effect of miRNA-137 targeting solute vector family member(SLC1A)5 on the neural function of rats with cerebral ischemia/reperfusion injury(CIRI)and its possible mechanism of regulating ferroptosis.Methods Sixty healthy adult SD rats were randomly divided into 6 groups:Sham group,Model group,miR-137 mimic group,miR-137 inhibitor group,miR-137 vector group,Ferrostatin 1 ferroptosis inhibitor(Fer-1)group(5 mg/kg),At 1 w before modeling,2μl viral vector was injected into the lateral ventricle of each virus group for 7 consecutive days;Longa method was used to evaluate the neurological function of rats on the 1st,3rd and 7th day modeling,and the samples were collected after the neurological function score on the 7th day,the expressions of reduced glutathione(GSH)and Ferric ion(Fe^(2+))were detected by biochemical kit.The expression levels of miR-137,SLC1A5,solute vector family 7 member(SLC7A)11 and glutathione peroxidase(GPX)4 in the brain tissues of each group were detected by qRT-polymerase chain reaction(PCR).Western blot was used to detect the protein expressions of SLC1A5,SLC7A11 and GPX4.The changes of iron death in mitochondria of neurons in each group were observed by electron microscopy.Results Compared with Sham group,neurological deficit score,Fe^(2+)content,SLC1A5 mRNA and protein expressions were significantly increased,GSH content,SLC7A11,GPX4 mRNA and protein expressions were significantly decreased in Model group(P<0.05,P<0.01).Mitochondrial shrinkage,mitochondrial membrane thickening and mitochondrial crest dissolution were observed by electron microscope.Compared with Model group,neurological deficit score,Fe^(2+)content,SLC1A5 mRNA and protein expressions were significantly decreased,GSH content,SLC7A11,GPX4 mRNA and protein expressions were significantly increased in miR-137 mimic group(P<0.05,P<0.01).Mitochondrial morphological damage was significantly improved,characterized by intact mitochondrial membrane,numerous mitochondrial cristae,and clear structure.Compared with

关 键 词：脑缺血再灌注损伤 miRNA-137 铁死亡 神经保护 

分 类 号：R245[医药卫生—针灸推拿学]