MicroRNA-29b inhibits TGF-β1-induced fibrosis via regulation of the TGF-β1/Smad pathway in primary human endometrial stromal cells.
MicroRNA-29b inhibits TGF-β1-induced fibrosis via regulation of the TGF-β1/Smad pathway in primary human endometrial stromal cells.
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DOI：
10.3892/mmr.2016.5062
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发表时间：
2016-05
影响因子：
3.4
通讯作者：
He Y
中科院分区：
文献类型：
--
作者：
Li J;Cen B;Chen S;He Y
转化生长因子（TGF）-β1在纤维化和器官功能障碍的调节中具有关键作用。在重要器官纤维化的发病和进展过程中，microRNA（miR）-29家族不规则地下调，外源性补充miR-29 b具有很强的抗纤维化能力。然而，TGF-β1是否能够引起子宫内膜纤维化，以及miR-29在子宫内膜纤维化中的作用尚不清楚。本研究采用RT-qPCR、免疫细胞化学、western blot分析、扫描电镜、免疫荧光染色、细胞增殖试验和流式细胞术等方法。结果显示，TGF-β1刺激后，子宫内膜间质细胞（ESCs）中1型胶原α 1（COL 1A 1）、α-平滑肌肌动蛋白（α-SMA）和磷酸化（p）-Smad 2/3的表达水平升高，而miR-29 b和母源性表达基因3（MEG 3）的表达水平降低，并呈时间和剂量依赖性。miR-29 b过表达可显著降低COL 1A 1和α-SMA的表达水平，并降低p-Smad 2/3的表达和核内积聚。此外，miR-29 b的异位过表达增加MEG 3的表达水平，抑制肌成纤维细胞样细胞增殖并诱导凋亡。这些发现表明，miR-29 b可能具有显著的抗纤维化作用，并且可能减轻TGF-β1诱导的ESC纤维化。因此，外源性miR-29 b可能成为治疗子宫内膜纤维化的潜在药物。
Transforming growth factor (TGF)-β1 has a key role in the regulation of fibrosis and organ dysfunction. During the pathogenesis and progression of vital organ fibrosis, the microRNA (miR)-29 family is irregularly downregulated and exogenous supplementation of miR-29b has a strong anti-fibrotic capacity. However, whether TGF-β1 is able to provoke endometrial fibrosis, and the role of miR-29 in endometrial fibrosis remain unclear. In the present study, RT-qPCR, immunocytochemistry, western blot analysis, scanning electron microscopy, immunofluorescence staining, cell proliferation assay and flow cytometric analysis were employed. The results demonstrated that the expression levels of collagen, type 1, alpha 1 (COL1A1), α-smooth muscle actin (α-SMA) and phosphorylated (p)-Smad2/3 were increased, whereas miR-29b and maternally expressed gene 3 (MEG3) were decreased in primary endometrial stromal cells (ESCs) in response to TGF-β1 stimulation, in a time and dose-dependent manner. Furthermore, overexpression of miR-29b markedly reduced the expression levels of COL1A1 and α-SMA, and decreased the expression and nuclear accumulation of p-Smad2/3. In addition, ectopic overexpression of miR-29b increased the expression levels of MEG3, inhibited myofibroblast-like cell proliferation and induced apoptosis. These findings indicated that miR-29b may have a significant anti-fibrotic role, and may attenuate TGF-β1-induced fibrosis in ESCs. Therefore, exogenous miR-29b may serve as a potential therapeutic agent for the treatment of endometrial fibrosis.