长链非编码 RNA MIAT 通过靶向 miR-200a 促进 BM-MSCs 向内皮细胞分化并恢复勃起功能障碍大鼠模型的勃起功能障碍。
LncRNA MIAT facilitated BM-MSCs differentiation into endothelial cells and restored erectile dysfunction via targeting miR-200a in a rat model of erectile dysfunction.
机构信息
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.
出版信息
Eur J Cell Biol. 2018 Apr;97(3):180-189. doi: 10.1016/j.ejcb.2018.02.001. Epub 2018 Feb 7.
BACKGROUND
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.
METHODS
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.
RESULTS
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.
CONCLUSION
LncRNA MIAT promoted BM-MSCs differentiation into ECs and restored ED via miR-200a.
背景
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。
方法
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。
结果
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。
结论
lncRNA MIAT 通过 miR-200a 促进 BM-MSCs 分化为 ECs 并恢复 ED。
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