Pirfenidone anti-fibrotic effects are partially mediated by the inhibition of MUC1 bioactivation.
Pirfenidone anti-fibrotic effects are partially mediated by the inhibition of MUC1 bioactivation.
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DOI：
10.18632/oncotarget.27526
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发表时间：
2020-04-14
期刊：
影响因子：
--
通讯作者：
Cortijo, Julio
中科院分区：
文献类型：
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作者：
Ballester, Beatriz;Milara, Javier;Cortijo, Julio
吡非尼酮是一种多效性分子，获批用于治疗特发性肺纤维化（IPF）。已证明吡非尼酮可下调转化生长因子-β 1（TGF-β 1）的细胞效应。然而，其抗纤维化的机制仍不清楚。在这里，我们的目的是分析吡非尼酮对TGF-β 1经典和非经典途径的影响，以及对最具特征性的IPF细胞过程的影响。本研究中观察到的结果表明，在肺泡A549和肺成纤维细胞MRC 5细胞中，TGF-β 1诱导的典型SMAD 3和非典型ERK 1/2磷酸化不受吡非尼酮抑制。相反，吡非尼酮抑制TGF-β 1诱导的MUC 1-CT Thr 41（1224）和Tyr 46（1229）磷酸化，从而降低β-连环蛋白激活。此外，ATII细胞和肺成纤维细胞中的免疫沉淀和免疫荧光研究表明，吡非尼酮抑制转录纤维化TGF-β 1诱导的磷酸化SMAD 3/MUC 1-CT/活性β-连环蛋白复合物的形成和核转位，从而抑制SMAD结合元件激活（SBE）。本研究还提供了吡非尼酮对TGF-β 1诱导的ATII向间充质细胞和成纤维细胞向肌成纤维细胞转化、成纤维细胞增殖以及ATII和成纤维细胞衰老的抑制作用的证据。因此，这表明吡非尼酮对TGF-β 1诱导的纤维化细胞过程的抑制作用是通过抑制MUC 1-CT磷酸化、β-连环蛋白活化、磷酸化SMAD 3/MUC 1-CT/活性β-连环蛋白的核复合物形成和SBE活化介导的，这可能对进一步开发抗纤维化IPF疗法具有价值。
Pirfenidone is a pleiotropic molecule approved to treat idiopathic pulmonary fibrosis (IPF). Pirfenidone has demonstrated to downregulate transforming growth factor-beta1 (TGF-beta1) cellular effects. However, its anti-fibrotic mechanism remains unclear. Here, we aim to analyze the effects of pirfenidone on the TGF-beta1 canonical and non-canonical pathways, as well as, on the most characteristic IPF cellular processes. Results observed in this work showed that TGF-beta1-induced canonical SMAD3 and non-canonical ERK1/2 phosphorylations were not inhibited by pirfenidone in alveolar A549 and lung fibroblasts MRC5 cells. In contrast, pirfenidone inhibited TGF-beta1-induced MUC1-CT Thr41 (1224) and Tyr46 (1229) phosphorylations, thus reducing the beta-catenin activation. Additionally, immunoprecipitation and immunofluorescence studies in ATII cells and lung fibroblasts showed that pirfenidone inhibited the formation and nuclear translocation of the transcriptional fibrotic TGF-beta1-induced phospho-SMAD3/MUC1-CT/active-beta-catenin complex, and consequently the SMAD-binding element activation (SBE). This study provided also evidence of the inhibitory effect of pirfenidone on the TGF-beta1-induced ATII to mesenchymal and fibroblast to myofibroblast transitions, fibroblast proliferation and ATII and fibroblast senescence. Therefore, it indicates that pirfenidone's inhibitory effect on TGF-beta1-induced fibrotic cellular processes is mediated by the inhibition of MUC1-CT phosphorylation, beta-catenin activation, nuclear complex formation of phospho-SMAD3/MUC1-CT/active beta-catenin and SBE activation, which may be of value to further develop anti-fibrotic IPF therapies.