Pirfenidone and nintedanib modulate properties of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis.
Pirfenidone and nintedanib modulate properties of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis.
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DOI：
10.1186/s12931-016-0328-5
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发表时间：
2016-02-04
影响因子：
5.8
通讯作者：
Kaarteenaho R
中科院分区：
文献类型：
--
作者：
Lehtonen ST;Veijola A;Karvonen H;Lappi-Blanco E;Sormunen R;Korpela S;Zagai U;Sköld MC;Kaarteenaho R
特发性肺纤维化（IPF）是一种无法治愈的肺部疾病，预后不良。成纤维细胞和肌成纤维细胞是纤维化过程中的关键细胞。最近，吡非尼酮和尼达尼布两种药物被批准用于临床，因为它们能够减缓疾病进展。这两种药物在体外细胞系统中的作用机制尚不清楚。因此，本研究的目的是检查吡非尼酮和尼达尼布对患有或不患有IPF患者的成纤维细胞和肌成纤维细胞结构和功能的影响。从对照肺（n = 4）或IPF（n = 7）收集并培养基质细胞。用吡非尼酮和/或尼达尼布处理细胞，通过Western分析和/或免疫电子显微镜测量细胞增殖、α平滑肌肌动蛋白（α-SMA）和纤连蛋白表达、透射电子显微镜测量超微结构特性以及胶原凝胶收缩和侵袭试验测量功能特性，评价处理效果。吡非尼酮和尼达尼布均以剂量依赖性方式降低成纤维细胞的体外增殖。1 mM吡非尼酮使对照肺的细胞数量减少至47%（p = 0.04），IPF细胞数量减少至42%（p = 0.04），1 μM尼达尼布相应地减少至67%（p = 0.04）和68%（p = 0.04）。如果两种药物一起使用，则观察到进一步减少的增殖。吡非尼酮和尼达尼布均能够减少α-SMA的量和肌纤维母细胞外观，尽管减少水平具有细胞系依赖性。在功能测定中，两种药物的作用也是可变的。我们得出结论，吡非尼酮和尼达尼布影响成纤维细胞和肌成纤维细胞的超微结构和功能。联合用药比单独用药更能减少细胞增殖。人肺源性细胞培养系统代表了筛选和测试纤维化疾病药物的潜在平台。本文的在线版本（doi：10.1186/s12931-016-0328-5）包含补充材料，可供授权用户使用。
Idiopathic pulmonary fibrosis (IPF) is an incurable lung disease with a poor prognosis. Fibroblasts and myofibroblasts are the key cells in the fibrotic process. Recently two drugs, pirfenidone and nintedanib, were approved for clinical use as they are able to slow down the disease progression. The mechanisms by which these two drugs act in in vitro cell systems are not known. The aim of this study was therefore to examine the effects of pirfenidone and nintedanib on fibroblasts and myofibroblasts structure and function established from patients with or without IPF. Stromal cells were collected and cultured from control lung (n = 4) or IPF (n = 7). The cells were treated with pirfenidone and/or nintedanib and the effect of treatment was evaluated by measuring cell proliferation, alpha smooth muscle actin (α-SMA) and fibronectin expression by Western analysis and/or immunoelectron microscopy, ultrastructural properties by transmission electron microscopy and functional properties by collagen gel contraction and invasion assays. Both pirfenidone and nintedanib reduced in vitro proliferation of fibroblastic cells in a dose dependent manner. The number of cells from control lung was reduced to 47 % (p = 0.04) and of IPF cells to 42 % (p = 0.04) by 1 mM pirfenidone and correspondingly to 67 % (p = 0.04) and 68 % (p = 0.04), by 1 μM nintedanib. If both drugs were used together, a further reduced proliferation was observed. Both pirfenidone and nintedanib were able to reduce the amount of α-SMA and the myofibroblastic appearance although the level of reduction was cell line dependent. In functional assays, the effect of both drugs was also variable. We conclude that the ultrastructure and function of fibroblasts and myofibroblasts are affected by pirfenidone and nintedanib. Combination of the drugs reduced cell proliferation more than either of them individually. Human lung derived cell culture systems represent a potential platform for screening and testing drugs for fibrotic diseases. The online version of this article (doi:10.1186/s12931-016-0328-5) contains supplementary material, which is available to authorized users.