miR-200b/c attenuates lipopolysaccharide-induced early pulmonary fibrosis by targeting ZEB1/2 via p38 MAPK and TGF-β/smad3 signaling pathways
miR-200b/c attenuates lipopolysaccharide-induced early pulmonary fibrosis by targeting ZEB1/2 via p38 MAPK and TGF-β/smad3 signaling pathways
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DOI：
10.1038/labinvest.2017.123
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发表时间：
2018-03-01
影响因子：
5
通讯作者：
Li, Yingchuan
中科院分区：
文献类型：
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作者：
Cao, Yongmei;Liu, Yujing;Li, Yingchuan
急性呼吸窘迫综合征（ARDS）早期引发的肺纤维化导致患者预后不良。然而，microRNA (miRNA) 是否可以作为 ARDS 期间早期肺纤维化的治疗靶点仍不清楚。在本研究中，我们评估了 miR-200 及其靶标 ZEB1/2 在肺组织中的作用和机制。通过脂多糖（LPS）三击方案建立ARDS引起的早期肺纤维化小鼠模型。在注射 LPS 前 1 天，将慢病毒包装的 miR-200b/c cDNA 或 ZEB1/2 shRNA 通过气管内注射到 C57BL/6 小鼠的肺部。在体外，用 miR-200b/c 或 SB203580/SIS3 预处理 30 分钟后，用 LPS 或 LPS + 转化生长因子-β (TGF-β) 刺激 RLE-6TN 细胞 24 小时。在肺纤维化的发展过程中，miR-200b/c和E-cadherin蛋白表达下降，而ZEB1/2 mRNA和蛋白以及波形蛋白和α-平滑肌肌动蛋白（α-SMA）蛋白水平逐渐升高。此外，miR-200b/c的过表达和ZEB1/2的沉默均显着减轻了肺部炎症和纤维化，减少了波形蛋白和α-SMA的表达，并增加了E-钙粘蛋白的水平。在 RLE-6TN 细胞中，LPS 与 TGF-β 联合发挥协同作用，增加波形蛋白和 α-SMA 蛋白水平，增加 p38 和 smad3 磷酸化，降低 E-cadherin 蛋白水平，而用 miR-200b/c 或 SB203580/SIS3 预处理可逆转这些作用。我们的研究结果表明，在 LPS 诱导的早期肺纤维化的发展过程中，miR-200b/c 下调，而 ZEB1/2 上调。 miR-200b/c通过靶向ZEB1/2发挥保护作用，这可能与抑制p38 MAPK和TGF-β/smad3信号通路有关。
Pulmonary fibrosis triggered during the early stage of acute respiratory distress syndrome (ARDS) contributes to poor prognosis in patients. However, whether microRNAs (miRNAs) can serve as therapeutic targets for early pulmonary fibrosis during ARDS is still largely unknown. In this study, we evaluated the effects and mechanisms of miR-200s and its targets ZEB1/2 in lung tissue. An early pulmonary fibrosis mouse model caused by ARDS was established via a lipopolysaccharide (LPS) three-hit regimen. Lentiviral packaged miR-200b/c cDNA or ZEB1/2 shRNA was intratracheally administered into the lungs of C57BL/6 mice 1 day before an LPS injection was administered. In vitro, following a 30-min pretreatment with miR-200b/c or SB203580/SIS3, RLE-6TN cells were stimulated by LPS or LPS + transforming growth factor-beta (TGF-beta) for 24 h. miR-200b/c and E-cadherin protein expression declined, whereas ZEB1/2 mRNA and protein and vimentin and alpha-smooth muscle actin (alpha-SMA) protein levels gradually increased during the development of pulmonary fibrosis. Furthermore, both the overexpression of miR-200b/c and the silencing of ZEB1/2 significantly alleviated pulmonary inflammation and fibrosis, reduced vimentin and alpha-SMA expression, and increased E-cadherin protein levels. In RLE-6TN cells, LPS combined with TGF-beta exerts synergistic effects of increasing vimentin and alpha-SMA protein levels, increasing p38 and smad3 phosphorylation and reducing E-cadherin protein levels, which were reversed by pretreatment with miR-200b/c or SB203580/SIS3. Our findings demonstrate that miR-200b/c was downregulated, whereas ZEB1/2 was upregulated in the development of LPS-induced early pulmonary fibrosis. miR-200b/c exerts a protective effect by targeting ZEB1/2, which may be associated with the inhibition of p38 MAPK and TGF-beta/smad3 signaling pathways.