MiR-150 Regulates Poststroke Cerebral Angiogenesis via Vascular Endothelial Growth Factor in Rats
MiR-150 Regulates Poststroke Cerebral Angiogenesis via Vascular Endothelial Growth Factor in Rats
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MiR-150 通过血管内皮生长因子调节大鼠中风后脑血管生成
DOI：
10.1111/cns.12525
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发表时间：
2016-06-01
影响因子：
5.5
通讯作者：
Hu, Bo
中科院分区：
文献类型：
--
作者：
He, Quan-Wei;Li, Qian;Hu, Bo
目的：血管生成是中风后恢复的统一目标。因此，探讨脑卒中后血管生成的机制具有重要意义。在这项研究中，我们报告了一种基于 miR-150 的脑卒中后血管生成机制。方法：建立大鼠大脑中动脉闭塞（MCAO）模型和氧糖剥夺细胞模型。分别通过FITC-葡聚糖测定、基质胶测定、Ki-67染色和伤口愈合测定来测量毛细血管密度、管形成、细胞增殖和细胞迁移。分别通过RT-PCR和Western blotting检测miR-150和血管内皮生长因子（VEGF）的表达。进行双荧光素酶测定以确认 miR-150 和 VEGF 之间的结合位点。结果：我们发现 miR-150 在脑缺血大鼠的大脑和血清中以及缺氧葡萄糖脑微血管内皮细胞 (BMVEC) 和星形胶质细胞中表达。上调 miR-150 表达可降低 MCAO 后大鼠梗塞边界区的血管密度，并减少 BMVEC 的管形成、增殖和迁移。我们还发现miR-150可以负向调节VEGF的表达，并且VEGF被证实是miR-150的直接靶标。此外，VEGF 介导 miR-150 对 BMVEC 管形成、增殖和迁移的功能。结论：我们的数据表明 miR-150 可以通过 VEGF 调节大鼠脑卒中后血管生成。
Aims: 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. Methods: 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. Results: 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. Conclusions: Our data suggested that miR-150 could regulate cerebral poststroke angiogenesis in rats through VEGF.