Upregulation of RGS2: a new mechanism for pirfenidone amelioration of pulmonary fibrosis.
Upregulation of RGS2: a new mechanism for pirfenidone amelioration of pulmonary fibrosis.
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DOI：
10.1186/s12931-016-0418-4
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发表时间：
2016-08-22
影响因子：
5.8
通讯作者：
Tu Y
中科院分区：
文献类型：
--
作者：
Xie Y;Jiang H;Zhang Q;Mehrotra S;Abel PW;Toews ML;Wolff DW;Rennard S;Panettieri RA Jr;Casale TB;Tu Y
吡非尼酮最近被批准用于治疗特发性肺纤维化。然而，吡非尼酮的治疗剂量非常高，导致副作用，限制了其剂量和治疗效果。了解吡非尼酮的分子作用机制可以提高其安全性和有效性。由于活化的成纤维细胞是与纤维化进展相关的关键效应细胞，本研究调查了肺成纤维细胞在吡非尼酮治疗后表达快速变化的基因，并探讨它们在吡非尼酮抗纤维化作用中的作用。我们使用基因芯片微阵列筛选在人肺成纤维细胞暴露于吡非尼酮后迅速上调的基因，并通过实时聚合酶链式反应和蛋白质印迹确认特定基因。在肺纤维化的细胞和动物模型上，通过生化和功能分析来确定它们的抗纤维化作用。我们发现G蛋白信号转导调节因子2(RGS2)是吡非尼酮诱导的早期基因。吡非尼酮治疗显著增加了人胎肺成纤维细胞系和分离自非特发性肺纤维化或特发性肺纤维化患者的原代肺成纤维细胞中RGS2的mRNA和蛋白表达。吡非尼酮处理或在人肺成纤维细胞中直接过表达重组RGS2可抑制凝血酶的促纤维化作用，而RGS2的缺失则加剧了博莱霉素诱导的小鼠肺纤维化和死亡率。在野生型RGS2基因敲除小鼠中，吡非尼酮治疗可减少博莱霉素诱导的肺纤维化，但不能。内源性RGS2具有抗纤维化作用。上调的RGS2对吡非尼酮的抗纤维化作用有重要作用。本文的在线版本(doi：10.1186/s12931-0160418-4)包含补充材料，授权用户可以使用。
Pirfenidone was recently approved for treatment of idiopathic pulmonary fibrosis. However, the therapeutic dose of pirfenidone is very high, causing side effects that limit its doses and therapeutic effectiveness. Understanding the molecular mechanisms of action of pirfenidone could improve its safety and efficacy. Because activated fibroblasts are critical effector cells associated with the progression of fibrosis, this study investigated the genes that change expression rapidly in response to pirfenidone treatment of pulmonary fibroblasts and explored their contributions to the anti-fibrotic effects of pirfenidone. We used the GeneChip microarray to screen for genes that were rapidly up-regulated upon exposure of human lung fibroblast cells to pirfenidone, with confirmation for specific genes by real-time PCR and western blots. Biochemical and functional analyses were used to establish their anti-fibrotic effects in cellular and animal models of pulmonary fibrosis. We identified Regulator of G-protein Signaling 2 (RGS2) as an early pirfenidone-induced gene. Treatment with pirfenidone significantly increased RGS2 mRNA and protein expression in both a human fetal lung fibroblast cell line and primary pulmonary fibroblasts isolated from patients without or with idiopathic pulmonary fibrosis. Pirfenidone treatment or direct overexpression of recombinant RGS2 in human lung fibroblasts inhibited the profibrotic effects of thrombin, whereas loss of RGS2 exacerbated bleomycin-induced pulmonary fibrosis and mortality in mice. Pirfenidone treatment reduced bleomycin-induced pulmonary fibrosis in wild-type but not RGS2 knockout mice. Endogenous RGS2 exhibits anti-fibrotic functions. Upregulated RGS2 contributes significantly to the anti-fibrotic effects of pirfenidone. The online version of this article (doi:10.1186/s12931-016-0418-4) contains supplementary material, which is available to authorized users.