Effect of MicroRNA-126a-3p on Bone Marrow Mesenchymal Stem Cells Repairing Blood-brain Barrier and Nerve Injury after Intracerebral Hemorrhage
Effect of MicroRNA-126a-3p on Bone Marrow Mesenchymal Stem Cells Repairing Blood-brain Barrier and Nerve Injury after Intracerebral Hemorrhage
复制标题
microRNA-126a-3p对骨髓间充质干细胞修复脑出血后血脑屏障及神经损伤的影响
DOI：
10.1016/j.jstrokecerebrovasdis.2020.104748
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发表时间：
2020-05-01
影响因子：
2.5
通讯作者：
Qian, Yuanyuan
中科院分区：
文献类型：
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作者：
Wang, Chunyan;Cao, Jingwei;Qian, Yuanyuan
目的：脑出血是一种严重威胁人类健康的疾病。在寻找更好的治疗脑出血的方法的同时，研究人员更关注的是寻找有效基因修饰干细胞的新技术。方法：采用体外分离、培养和鉴定骨髓间充质干细胞（MSCs）。进一步，将转染表达microRNA-126a-3p （miR-126）的慢病毒的MSCs注射到该型中。目的探讨胶原酶诱导脑出血大鼠血脑屏障（BBB）和神经损伤在体内的恢复作用。结果：MSCs表面标记分子（CD29: 98.5%; CD90: 96.5%）高表达，血细胞表面标记分子（CD45: 2%）负表达。同时，我们证实miR-126促进了MSCs向血管内皮细胞的分化，这是由于标记物（CD31和VE-cadherin）的增加。移植mir -126修饰的MSCs后，改良的神经系统严重程度评分、改良的肢体放置测试评分、脑含水量和evans蓝含量均降低。体内免疫组化染色发现miR-126加速了MSCs向血管内皮细胞的分化。HE染色显示，与ICH +载体- mscs组相比，水肿面积明显减少。TUNEL实验表明，mir -126修饰的MSCs可减轻脑组织细胞凋亡。此外，ICH+miR-126-MSCs组蛋白酶活化受体-1和基质金属蛋白酶-9的mRNA和蛋白表达减少，而封闭带-1 （ZO-1）和claudin-5的表达增强。免疫荧光分析显示，mir -126修饰的MSCs减少了紧密连接的破坏（ZO-1和claudin-5）。结论：所有数据表明，mir -126修饰的MSCs可修复脑出血后血脑屏障和神经损伤。
Objective: Intracerebral hemorrhage (ICH) is a disease that threatens human health due to its high morbidity and mortality. On behalf of finding the better methods in the treatment of ICH, researchers pay more attention to a new technology which is finding effective genes to modify stem cells. Methods: In this study, we isolated, cultured and identified bone marrow mesenchymal stem cells (MSCs) in vitro. Further, the MSCs (transfected with lentivirus expressing microRNA-126a-3p (miR-126)) were injected into the type. collagenase-induced ICH rats to investigate the recovery effects of blood-brain barrier (BBB) and nerve damage in vivo. Results: The MSCs surface marker molecules (CD29: 98.5%; CD90: 96.5%) were highly expressed, and the blood cell surface molecule was negatively expressed (CD45: 2%). Meanwhile, it was verified that miR-126 facilitated the differentiation of MSCs into vascular endothelial cells, owing to the rise of markers (CD31 and VE-cadherin). The modified neurological severity score, modified limb placing test score, brain water content and evans blue content were reduced after transplanted miR-126-modified MSCs. It was found that miR-126 accelerated the differentiation of MSCs into vascular endothelial cells via immunohistochemical staining in vivo. HE staining indicated the area of edema was obviously decreased compared with that in ICH + vector-MSCs group. MiR-126-modified MSCs alleviated the cell apoptosis in brain tissues by TUNEL assay. In addition, the mRNA and protein expression of protease activated receptor-1 and matrix metalloproteinase-9 were diminished, whilst the expression of zonula occludens-1 (ZO-1) and claudin-5 were enhanced in ICH+miR-126-MSCs group. Immunofluorescence assay revealed that miR-126-modified MSCs decreased the disruption of tight junction (ZO-1 and claudin-5). Conclusions: All data illustrate that miR-126-modified MSCs repair BBB and nerve injury after ICH.