Wang Chunyan, Cao Jingwei, Duan Shurong, Xu Ran, Yu Hongli, Huo Xin, Qian Yuanyuan
Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, People's Republic of China.
Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, People's Republic of China.
J Stroke Cerebrovasc Dis. 2020 May;29(5):104748. doi: 10.1016/j.jstrokecerebrovasdis.2020.104748. Epub 2020 Mar 9.
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.
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 Ⅶ collagenase-induced ICH rats to investigate the recovery effects of blood-brain barrier (BBB) and nerve damage in vivo.
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).
All data illustrate that miR-126-modified MSCs repair BBB and nerve injury after ICH.
脑出血（ICH）因其高发病率和死亡率而成为威胁人类健康的疾病。为了找到更好的脑出血治疗方法，研究人员更加关注一种新技术，即寻找有效的基因来修饰干细胞。
在本研究中，我们在体外分离、培养并鉴定了骨髓间充质干细胞（MSCs）。进一步地，将转染了表达微小RNA-126a-3p（miR-126）的慢病毒的MSCs注射到Ⅶ型胶原酶诱导的脑出血大鼠体内，以研究其对血脑屏障（BBB）和神经损伤的体内修复作用。
MSCs表面标记分子（CD29：98.5%；CD90：96.5%）高表达，血细胞表面分子呈阴性表达（CD45：2%）。同时，由于标记物（CD31和VE-钙黏蛋白）的增加，证实miR-126促进了MSCs向血管内皮细胞的分化。移植miR-126修饰的MSCs后，改良神经功能缺损评分、改良肢体放置试验评分、脑含水量和伊文思蓝含量均降低。通过体内免疫组织化学染色发现，miR-126加速了MSCs向血管内皮细胞的分化。苏木精-伊红（HE）染色表明，与脑出血+载体-MSCs组相比，水肿面积明显减小。通过末端脱氧核苷酸转移酶介导的缺口末端标记法（TUNEL）检测发现，miR-修饰的MSCs减轻了脑组织中的细胞凋亡。此外，在脑出血+miR-126-MSCs组中，蛋白酶激活受体-1和基质金属蛋白酶-9的mRNA和蛋白表达降低，而紧密连接蛋白-1（ZO-1）和闭合蛋白-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（ZO-1和闭合蛋白-5）的破坏。
所有数据表明，miR-126修饰的MSCs可修复脑出血后的血脑屏障和神经损伤。
