Department of Urology and Andrology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, PR China.
Department of Urology and Andrology, Zhongnan Hospital, Wuhan University, Wuhan 430071, PR China.
Eur J Cell Biol. 2018 Apr;97(3):180-189. doi: 10.1016/j.ejcb.2018.02.001. Epub 2018 Feb 7.
Bone-marrow derived mesenchymal stem cells (BM-MSCs) implantation effectively restored rats' erectile dysfunction (ED). Long noncoding RNA (LncRNA)-myocardial infarction-associated transcript (MIAT) has been reported to play an important role in regulating endothelial cells (ECs) function via vascular endothelial growth factor (VEGF) that induced BM-MSCs differentiation into ECs. However, the molecular functions and biological roles of lncRNA MIAT in ED remained unclear.
The rat model of ED was established. Quantitative real-time PCR (qRT-PCR) and western blotting were used to detect the expression of lncRNA MIAT, von Willebrand factor (vWF), vascular endothelial cadherin (VE-cadherin), endothelial NO synthase (eNOS) and VEGF following BM-MSCs transfection. Erectile function was evaluated by intra-cavernous pressure/mean artery pressure (ICP/MAP). Furthermore, RNA immunoprecipitation (RIP) assay and RNA pull down as well as luciferase reporter assay were carried out to examine the interaction among lncRNA MIAT, miR-200a and VEGF.
BM-MSCs restored ED by upregulating lncRNA MIAT. LncRNA MIAT was upregulated in a time-dependent manner during BM-MSCs differentiation into ECs. LncRNA MIAT regulated VEGF via targeting miR-200a, thereby promoting BM-MSCs differentiation into ECs. LncRNA MIAT knockdown in vivo abolished the effect of BM-MSCs on ED.
LncRNA MIAT promoted BM-MSCs differentiation into ECs and restored ED via miR-200a.
骨髓间充质干细胞（BM-MSCs）移植有效地恢复了大鼠的勃起功能障碍（ED）。长链非编码 RNA（lncRNA）-心肌梗塞相关转录物（MIAT）已被报道通过血管内皮生长因子（VEGF）在调节内皮细胞（ECs）功能方面发挥重要作用，从而诱导 BM-MSCs 分化为 ECs。然而，lncRNA MIAT 在 ED 中的分子功能和生物学作用仍不清楚。
建立 ED 大鼠模型。采用定量实时 PCR（qRT-PCR）和 Western blot 检测 BM-MSCs 转染后 lncRNA MIAT、血管性血友病因子（vWF）、血管内皮钙黏蛋白（VE-cadherin）、内皮型一氧化氮合酶（eNOS）和 VEGF 的表达。通过阴茎海绵体内压/平均动脉压（ICP/MAP）评估勃起功能。此外，还进行了 RNA 免疫沉淀（RIP）试验、RNA 下拉和荧光素酶报告基因检测，以检验 lncRNA MIAT、miR-200a 和 VEGF 之间的相互作用。
BM-MSCs 通过上调 lncRNA MIAT 恢复 ED。在 BM-MSCs 分化为 ECs 的过程中，lncRNA MIAT 呈时间依赖性上调。lncRNA MIAT 通过靶向 miR-200a 调节 VEGF，从而促进 BM-MSCs 分化为 ECs。体内 lncRNA MIAT 敲低消除了 BM-MSCs 对 ED 的作用。
lncRNA MIAT 通过 miR-200a 促进 BM-MSCs 分化为 ECs 并恢复 ED。
