Department of Neurology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
CNS Neurosci Ther. 2016 Jun;22(6):507-17. doi: 10.1111/cns.12525. Epub 2016 Feb 28.
Angiogenesis is a harmonized target for poststroke recovery. Therefore, exploring the mechanisms involved in angiogenesis after stroke is vitally significant. In this study, we are reporting a miR-150-based mechanism underlying cerebral poststroke angiogenesis.
Rat models of middle cerebral artery occlusion (MCAO) and cell models of oxygen-glucose deprivation were conducted. Capillary density, tube formation, cell proliferation, and cell migration were measured by FITC-dextran assay, matrigel assay, Ki-67 staining, and wound healing assay, respectively. The expression of miR-150 and vascular endothelial growth factor (VEGF) was, respectively, measured by RT-PCR and Western blotting. Dual-luciferase assay was conducted to confirm the binding sites between miR-150 and VEGF.
We found that miR-150 expression in the brain and serum of rats subjected to cerebral ischemia, and in oxygen-glucose-deprived brain microvascular endothelial cells (BMVECs) and astrocytes. Upregulation of miR-150 expression could decrease vascular density of infarct border zone in rat after MCAO and decrease tube formation, proliferation, and migration of BMVECs. We also found that miR-150 could negatively regulate the expression of VEGF, and VEGF was confirmed to be a direct target of miR-150. Moreover, VEGF mediated the function of miR-150 on tube formation, proliferation, and migration of BMVECs.
Our data suggested that miR-150 could regulate cerebral poststroke angiogenesis in rats through VEGF.
血管生成是脑卒中后恢复的一个协调靶点。因此，探索脑卒中后血管生成的相关机制至关重要。本研究报告了一个基于 miR-150 的脑卒中后脑内血管生成的机制。
建立大脑中动脉闭塞（MCAO）大鼠模型和氧葡萄糖剥夺细胞模型。通过 FITC-葡聚糖测定法、基质胶测定法、Ki-67 染色法和划痕愈合试验分别测量毛细血管密度、管形成、细胞增殖和细胞迁移。分别通过 RT-PCR 和 Western blot 测定 miR-150 和血管内皮生长因子（VEGF）的表达。通过双荧光素酶报告基因实验证实 miR-150 与 VEGF 之间的结合位点。
我们发现，脑缺血大鼠大脑和血清中 miR-150 表达增加，氧葡萄糖剥夺的脑微血管内皮细胞（BMVECs）和星形胶质细胞中 miR-150 表达也增加。上调 miR-150 表达可减少 MCAO 后大鼠梗死边缘区的血管密度，并减少 BMVECs 的管形成、增殖和迁移。我们还发现，miR-150 可负向调节 VEGF 的表达，而 VEGF 被确认为 miR-150 的直接靶标。此外，VEGF 介导了 miR-150 对 BMVECs 管形成、增殖和迁移的功能。
我们的数据表明，miR-150 可通过 VEGF 调节大鼠脑卒中后血管生成。
