Lentivirus-mediated microRNA-124 gene-modified bone marrow mesenchymal stem cell transplantation promotes the repair of spinal cord injury in rats.
Lentivirus-mediated microRNA-124 gene-modified bone marrow mesenchymal stem cell transplantation promotes the repair of spinal cord injury in rats.
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DOI：
10.1038/emm.2017.48
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发表时间：
2017-05-19
影响因子：
12.8
通讯作者：
Fan CY
中科院分区：
文献类型：
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作者：
Song JL;Zheng W;Chen W;Qian Y;Ouyang YM;Fan CY
我们的研究旨在探讨慢病毒介导的微小RNA-124（miR-124）基因修饰的骨髓间充质干细胞（BMSC）移植对大鼠脊髓损伤（SCI）修复的影响。 BMSCs是从大鼠的骨髓中分离出来的。使用荧光素酶报告基因测定法鉴定了靶基因 miR-124。选取72只大鼠进行SCI模型构建，随机分为空白组、假手术组、SCI组、阴性对照（NC）组、过表达miR-124组和si-PDXK组。采用实时定量聚合酶链反应和蛋白质印迹法检测miR-124 mRNA表达以及吡哆醛激酶（PDXK）mRNA和蛋白表达。使用 Basso、Beattie 和 Bresnahan (BBB) 量表评估大鼠的运动能力。使用免疫组织化学检测 Brdu、神经元特异性烯醇化酶 (NSE)、神经丝 (NF) 和微管相关蛋白 2 (MAP2)。使用酶联免疫吸附测定法测量促甲状腺素释放激素（TRH）、前列环素（PGI2）和神经节苷脂（GM）的表达水平。 PDXK被确定为miR-124的靶基因。过表达的miR-124组比SCI、NC和si-PDXK组表现出更高的miR-124表达。与SCI组和NC组相比，过表达miR-124和si-PDXK组PDXK表达下调，移植后7、21和35天BBB评分显着升高。过表达miR-124组双标阳性细胞密度（Brdu+NSE/NF/MAP2）及TRH、PGI2、GM表达水平显着高于NC组和SCI组。这些结果表明miR-124靶向PDXK加速BMSCs向神经细胞的分化并促进SCI修复。
Our study aims to explore the effects of lentivirus-mediated microRNA-124 (miR-124) gene-modified bone marrow mesenchymal stem cell (BMSC) transplantation on the repair of spinal cord injury (SCI) in rats. BMSCs were isolated from the bone marrow of rats. The target gene miR-124 was identified using a luciferase-reporter gene assay. Seventy-two rats were selected for construction of the SCI model, and the rats were randomly divided into the blank group, sham group, SCI group, negative control (NC) group, overexpressed miR-124 group and si-PDXK group. The mRNA expression of miR-124 and the mRNA and protein expression of pyridoxal kinase (PDXK) were detected by quantitative real-time polymerase chain reaction and western blotting. The locomotor capacity of the rats was evaluated using the Basso, Beattie and Bresnahan (BBB) scale. Brdu, neuron-specific enolase (NSE), neurofilament (NF) and microtubule-associated protein 2 (MAP2) were detected using immunohistochemistry. The expression levels of thyrotropin-releasing hormone (TRH), prostacyclin (PGI2) and gangliosides (GM) were measured using an enzyme-linked immunosorbent assay. PDXK was identified as the target gene of miR-124. The overexpressed miR-124 group exhibited higher miR-124 expression than the SCI, NC and si-PDXK groups. Compared with the SCI and NC groups, the PDXK expression was downregulated in the overexpressed miR-124 and si-PDXK groups, and the BBB scores were significantly increased 7, 21 and 35 days after transplantation. The double-labeled positive cell densities (Brdu+NSE/NF/MAP2) and the expression levels of TRH, PGI2 and GM in the overexpressed miR-124 group were significantly higher than those in the NC and SCI groups. These results indicated that miR-124 targeted PDXK to accelerate the differentiation of BMSCs into neurocytes and promote SCI repair.