Department of Neurology, First Affiliated Hospital of Anhui Medical University, Hefei, PR China.
Department of Neurology, First Affiliated Hospital of Anhui Medical University, Hefei, PR China.
Kaohsiung J Med Sci. 2017 Aug;33(8):369-378. doi: 10.1016/j.kjms.2017.05.010. Epub 2017 Jun 9.
The study aims to explore how microRNA-146a/b (miR-146a/b) regulates the function of endothelial progenitor cells (EPCs) in acute ischemic stroke in mice. Eighty male SPF C57BL/6J mice were evenly divided into the model-6 h, model-12 h, model-24 h (mice suffered from middle cerebral artery occlusion [MCAO] for 6 h, 12 h and model-24 h) and normal groups. EPCs were transfected and assigned into the control, MCAO, MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups. The qRT-PCR was used to detect miR-146a/b expression in EPCs. Expressions of tumor necrosis factor receptor-associated factor 6 (TRAF6) and interleukin-1 receptor-associated kinase 1 (IRAK1) were detected using western blotting. Cell proliferation and migration of EPCs were testified using CCK-8 assay and scratch test, respectively. Angiogenesis ability of EPCs was observed under microscope. MiR-146a and miR-146b expressions were lower in the model groups than the normal group. There were up-regulated TRAF6 and IRAK1 expressions in the model-6 h, model-12 h and model-24 h groups compared with the normal group. And there were down-regulated TRAF6 and IRAK1 expressions in the MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups than in the MCAO group. Compared with the control group, the proliferation, migration and angiogenesis ability of EPCs were significantly lower in the MCAO group, but higher in the MCAO-miR-146a, MCAO-miR-146b and MCAO-miR-146a/b groups. Besides, the miR-146a/b group showed more enhancement than the MCAO-miR-146a and MCAO-miR-146b groups. MiR-146a/b could down-regulate the TRAF6 and IRAK1 expressions and promote proliferation, migration and angiogenesis ability of EPCs, which was important for recovery of patients with hyperacute ischemic stroke.
该研究旨在探讨 microRNA-146a/b(miR-146a/b)如何调节急性缺血性脑卒中小鼠内皮祖细胞(EPCs)的功能。80 只雄性 SPF C57BL/6J 小鼠平均分为模型-6h、模型-12h、模型-24h(大脑中动脉闭塞[MCAO] 6h、12h 和模型-24h)和正常组。转染 EPCs 并分为对照组、MCAO 组、MCAO-miR-146a 组、MCAO-miR-146b 组和 MCAO-miR-146a/b 组。qRT-PCR 检测 EPCs 中 miR-146a/b 的表达。Western blot 检测肿瘤坏死因子受体相关因子 6(TRAF6)和白细胞介素 1 受体相关激酶 1(IRAK1)的表达。CCK-8 检测 EPC 增殖,划痕试验检测 EPC 迁移,观察 EPC 血管生成能力。与正常组相比,模型组 miR-146a 和 miR-146b 的表达水平较低。与正常组相比,模型-6h、模型-12h 和模型-24h 组 TRAF6 和 IRAK1 的表达水平升高。MCAO-miR-146a、MCAO-miR-146b 和 MCAO-miR-146a/b 组 TRAF6 和 IRAK1 的表达水平低于 MCAO 组。与对照组相比,MCAO 组 EPC 的增殖、迁移和血管生成能力明显降低,而 MCAO-miR-146a、MCAO-miR-146b 和 MCAO-miR-146a/b 组则明显升高。此外,miR-146a/b 组的增强作用大于 MCAO-miR-146a 和 MCAO-miR-146b 组。miR-146a/b 可下调 TRAF6 和 IRAK1 的表达,促进 EPC 的增殖、迁移和血管生成能力,这对超急性缺血性脑卒中患者的恢复至关重要。