Department of Geriatrics, Weifang People's Hospital, Weifang, Shandong 261041, P.R. China.
Mol Med Rep. 2018 Apr;17(4):5300-5305. doi: 10.3892/mmr.2018.8513. Epub 2018 Jan 29.
Stroke is the most common cause of mortality worldwide. Post-stroke angiogenesis is of great significance to the treatment of strokes. The aim of the present study was to investigate the mechanism underlying the angiogenesis-promoting effect of microRNA‑126 (miR‑126)‑associated signaling pathways using a stroke model in vivo and a cell migration model in vitro. Bone marrow‑derived endothelial progenitor cells (EPCs) were extracted and identified using a density gradient method. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was performed to examine the expression levels of miR‑126 and C‑X‑C chemokine receptor type 7 (CXCR7). Target genes of miR‑126 were analyzed using TargetScan software version 7.1 (www.targetscan.org/). In addition, a reporter gene assay and RT‑qPCR were performed to determine the target genes of miR‑126. The effect of miR‑126 on cell migration was examined using a cell migration model in vitro and a middle cerebral artery occlusion model of mice was established in vivo. The miR‑126 antagomir‑treated EPCs were infused into stroke mice. Microvessel density, nerve function score and infarction volume were assessed. Flow cytometric analysis indicated that cluster of differentiation (CD)34, CD133 and vascular endothelial growth factor receptor 2 were partly expressed on the cell surface of bone marrow‑derived EPCs. In addition, the expression levels of Di‑acetylated‑low density lipoprotein and Ulex europaeus agglutinin 1 were positive. Stromal cell‑derived factor 1 (SDF-1) was identified as a target gene of miR‑126, which was confirmed by a reporter gene assay and RT‑qPCR. Cell migration examination demonstrated that the neutralizing antibody of CXCR7 blocked miR‑126 angomir‑induced migration of EPCs. Microvessel density increased, while nerve function score and infarction volume decreased following infusion of miR-126 angomir‑treated EPCs. Furthermore, miR‑126 angomir improved the efficacy of EPC treatment. Thus, miR‑126 improved the migration of EPCs via the miR‑126/SDF‑1/CXCR7 signaling pathway.
中风是全球最常见的死亡原因。中风后血管生成对中风的治疗具有重要意义。本研究旨在通过体内中风模型和体外细胞迁移模型研究微小 RNA-126 (miR-126）相关信号通路促进血管生成的机制。采用密度梯度法提取并鉴定骨髓源性内皮祖细胞(EPCs)。采用逆转录-定量聚合酶链反应(RT-qPCR)检测 miR-126 和 C-X-C 趋化因子受体 7(CXCR7)的表达水平。使用 TargetScan 软件版本 7.1(www.targetscan.org/)分析 miR-126 的靶基因。此外，进行报告基因检测和 RT-qPCR 以确定 miR-126 的靶基因。采用体外细胞迁移模型和体内大脑中动脉闭塞模型研究 miR-126 对细胞迁移的影响。将 miR-126 反义寡核苷酸处理的 EPCs 注入中风小鼠体内。评估微血管密度、神经功能评分和梗死体积。流式细胞术分析表明，骨髓源性 EPCs 的细胞表面部分表达分化抗原（CD）34、CD133 和血管内皮生长因子受体 2。此外，二乙酰化低密度脂蛋白和荆豆凝集素 1 的表达呈阳性。基质细胞衍生因子 1(SDF-1)被鉴定为 miR-126 的靶基因，这通过报告基因检测和 RT-qPCR 得到证实。细胞迁移检测表明，CXCR7 的中和抗体阻断了 miR-126 angomir 诱导的 EPC 迁移。miR-126 angomir 处理的 EPCs 输注后，微血管密度增加，神经功能评分和梗死体积降低。此外，miR-126 angomir 改善了 EPC 治疗的效果。因此，miR-126 通过 miR-126/SDF-1/CXCR7 信号通路改善 EPC 的迁移。
