蛛网膜下腔出血后外泌体源性miR-630对微血管内皮细胞功能的影响  

Effects of exosomal miR-630 on brain microvascular endothelial cells’function after subarachnoid hemorrhage

作　　者：孙雷涛[1] 杜德勇[1] 李勐[1] 李泽福[1] 张文生[1] SUN Leitao;DU Deyong;LI Meng;LI Zefu;ZHANG Wensheng(Department of Neurosurgery,Binzhou Medical University Hospital,Binzhou 256603,China)

机构地区：[1]滨州医学院附属医院神经外科,山东滨州256603

出　　处：《西安交通大学学报（医学版）》2021年第5期645-650,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：山东省医药卫生科技发展计划项目(No.2017WS367,2018WS550)。

摘　　要：目的探讨蛛网膜下腔出血后脑脊液外泌体源性miR-630与脑微血管内皮细胞(BMECs)功能之间的关系。方法建立蛛网膜下腔出血内皮细胞模型,通过MTT和细胞周期分析,评估血性脑脊液(BCSF)对BMECs增殖的影响。采用qRT-PCR和免疫荧光染色法检测内皮细胞紧密连接蛋白(ZO-1)和黏附分子(ICAM-1、VCAM-1)的表达变化。放射免疫法检测NOX浓度变化。分离并鉴定实验组(BCSF共孵育)与对照组(正常脑脊液)中脑脊液外泌体,比较两组脑脊液外泌体源性miR-630的表达差异。观察应用miR-630 mimics干预后BMECs的功能变化。结果实验组BMECs增殖明显抑制,ICAM-1、VCAM-1和ZO-1的mRNA及蛋白表达均明显降低,NOX浓度明显降低(P<0.05),内皮细胞功能受到明显抑制。成功分离并鉴定脑脊液外泌体,实验组外泌体miR-630的表达显著低于对照组(P<0.05);外源性miR-630 mimics干预后BMECs功能有了显著改善。结论蛛网膜下腔出血后脑脊液外泌体miR-630的低表达与BMECs损伤关系密切。Objective To investigate the relationship between cerebrospinal fluid exosome-derived miR-630 and the function of brain microvascular endothelial cells(BMECs).Methods The subarachnoid hemorrhage endothelial cell model was established to evaluate the effect of hemorrhagic cerebrospinal fluid(BCSF)on BMECs’proliferation by MTT assay and cell cycle analysis.qRT-PCR and immunofluorescence staining were used to detect the expressions of endothelial cell tight junction protein(ZO-1)and adhesion molecule(ICAM-1 and VCAM-1).Changes in NOx concentration were detected by radioimmunoassay.The cerebrospinal fluid exosomes in the experimental group(co-incubated with BCSF)and the control group(normal cerebrospinal fluid)were isolated and identified,and differences in the expressions of cerebrospinal fluid exosomal miR-630 between the two groups were compared.BMECs work changes after the intervention with miR-630 analogue were observed.Results The proliferation of BMECs was significantly inhibited in the experimental group;the mRNA and protein levels of ICAM-1,VCAM-1 and ZO-1 were significantly decreased,and the function of endothelial cells was significantly inhibited(P<0.05).After the successful separation and identification of cerebrospinal fluid exosomes,the expression of miR-630 was significantly lower in the experimental group than in the control group(P<0.05).The function of BMECs was significantly improved with miR-630 mimics.Conclusion The low expression of miR-630 in cerebrospinal fluid exosomes after subarachnoid hemorrhage is closely related to BMECs injury.

关 键 词：蛛网膜下腔出血 外泌体 脑微血管内皮细胞 miR-630 

分 类 号：R743.35[医药卫生—神经病学与精神病学]