miR-451/MIF信号通路在脑出血后血脑屏障破坏中的作用  

The role of miR-451/MIF signaling pathway in the disruption of blood-brain barrier after intracerebral hemorrhage

作　　者：白霜 陈施玲 张格 刘霞 陈丹阳 唐洲平[1] 唐颖馨 Bai Shuang;Chen Shiling;Zhang Ge(Department of Neurology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030)

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《卒中与神经疾病》2022年第4期305-310,共6页Stroke and Nervous Diseases

基　　金：国家自然科学基金(编号为81873750、82071330、92148206)。

摘　　要：目的观察脑出血后微小RNA-451(MicroRNA,miR-451)/巨噬细胞迁移抑制因子(Macrophage migrationinhibitory factor,MIF)信号通路的变化及其对血脑屏障紧密连接的影响。方法体外培养人脑微血管内皮细胞(Human brain microvascular endothelial cells,hBMECs),随机分为对照组和血红素组;对照组细胞给予常规培养基,血红素组细胞的培养基中加入60μM血红素分别培养12、24和48 h,实时荧光定量聚合酶链反应(Polymerase chain reaction,PCR)检测各组各时间点miR-451转录水平;体外培养hBMECs细胞,随机分为空白组、血红素组、阴性模拟物组和miR-451模拟物组,培养24 h;酶联免疫吸附法(Enzyme linked immunosorbent assay,ELASA)检测各组培养基上清中MIF分泌水平,实时荧光定量PCR检测各组MIF mRNA的转录水平及紧密连接相关蛋白[咬合蛋白Occludin、闭合蛋白Claudin和闭合小环蛋白1(Zonula occluden-1,ZO-1)]mRNA的转录水平。结果hBMECs细胞培养基中加入60μM血红素分别作用12、24和48 h后各时间点的miR-451转录水平均低于同时间点的对照组(P均<0.05),且24 h时miR-451转录水平最低。各组hBMECs细胞培养24 h后血红素组培养基上清中的MIF蛋白的水平和细胞内MIF mRNA的转录水平均高于空白组(P均<0.05);miR-451模拟物组上清中的MIF蛋白水平和细胞内MIF mRNA的转录水平均低于阴性模拟物组(P均<0.05)。各组hBMECs细胞培养24 h后血红素组的细胞内Occludin mRNA,Claudin-5 mRNA和Zo-1 mRNA的转录水平低于空白组(P<0.05),miR-451模拟物组细胞内Occludin mRNA,Claudin-5 mRNA和Zo-1 mRNA的转录水平均高于阴性模拟物组(P<0.05)。结论miR-451/MIF信号通路可能参与了脑出血后血脑屏障紧密连接的破坏,给予外源性补充miR-451模拟物可能有助于维持血脑屏障的完整性。Objective To investigate the role of microRNA-451(miR-451)/macrophage migration inhibitory factor(MIF)signal pathway in tight junctions of blood-brain barrier(BBB)after cerebral hemorrhages.Methods Human brain microvascular endothelial cells(hBMECs)were cultured and randomly divided into the control group and the hemin group.hBMECs were cultured with normal medium or medium with 60Μm hemin for 12 h,24 h,or 48 h.Transcriptional levels of miR-451 were detected by real-time PCR(RT-PCR).hBMECs were cultured for 24 h and randomly divided into the blank control group,hemin group,NC mimic group,and miR-451 mimic group.MIF protein in the supernatant of each group was detected by ELISA and transcriptional levels of MIF mRNA,OccludinmRNA,Claudin-5,and ZO-1 mRNA were detected by RT-PCR.Results Transcriptional levels of miR-451 in the hemin group were lower than those in the control group after 12 h,24 h,and 48 h treatment with hemin(all P<0.05).The MIF level in the supernatant of hemin group was higher than that in the control group(P<0.05),and the transcription level of MIF mRNA in hBMECs of hemin group was higher than that of the control group(P<0.05).The MIF protein in the supernatant of miR-451 mimic group was lower than that in the NC mimic group(P<0.05).The transcription level of MIF mRNA in hBMECs of miR-451 mimic group was lower than that in the NC mimic group(P<0.05).The transcription levels of Occlud in mRNA,Claudin-5 mRNA,and Zo-1 mRNA of hBMECs in the hemin group were lower than those in the control group(all P<0.05).The transcription levels of Occlud in mRNA,Claudin-5 mRNA and Zo-1 mRNA of hBMECs in miR-451 mimic group were higher than those in NC mimic group(all P<0.05).Conclusion The miR-451/MIF signaling pathway may be involved in the destruction of tight junctions of BBB after intracerebral hemorrhage.Exogenous supplementation of miR-451 mimics may help maintain the integrity of the BBB.

关 键 词：微小RNA 巨噬细胞迁移抑制因子 脑出血 血脑屏障 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学] R743.34