Pirfenidone attenuates bleomycin-induced pulmonary fibrosis in mice by regulating Nrf2/Bach1 equilibrium.
Pirfenidone attenuates bleomycin-induced pulmonary fibrosis in mice by regulating Nrf2/Bach1 equilibrium.
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DOI：
10.1186/s12890-017-0405-7
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发表时间：
2017-04-18
影响因子：
3.1
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Liu Y;Lu F;Kang L;Wang Z;Wang Y
氧化应激是参与特发性肺纤维化（IPF）发病的重要因素之一。核因子-红细胞相关因子2 (Nrf2)/[BTB (broad-complex, tramtrack and brick -a-brac)]和CNC （cap 'n 'collar protein）同源性1，Bach1]的平衡决定了抗氧化因子的表达水平，进而调节氧化/抗氧化能力的功能。吡非尼酮（PFD）是目前批准用于IPF治疗的两种药物之一。PFD调节细胞内抗氧化剂，抑制炎症细胞因子的分泌和胶原合成。然而，其抗氧化作用的机制尚不清楚。研究PFD对转化生长因子β1 （TGF-β1）诱导小鼠肺成纤维细胞（MLF）和博来霉素诱导小鼠肺纤维化的影响。采用RT-PCR和Western blot检测氧化应激相关因子Nrf2/Bach1及其下游抗氧化因子血红素加氧酶-1 （Ho-1）和谷胱甘肽过氧化物酶1 (Gpx1) mRNA和蛋白水平。ELISA法检测MLF上清液、血清、支气管肺泡灌洗液（BALF）中纤维化相关细胞因子白介素-6 （IL-6）、肌成纤维细胞标志物1型胶原α1 （COL1A1）水平以及血清和BALF中丙二醛（MDA）水平；采用2′，7′-双乙酸二氯荧光素（DCFH-DA）法测定小鼠活性氧（ROS）生成，HE和Masson法观察小鼠肺病理形态学变化，探讨博来霉素抗氧化机制及对肺纤维化的治疗作用。PFD抑制TGF-β1诱导的小鼠肺成纤维细胞和博来霉素诱导的肺纤维化组织中Bach1 mRNA和蛋白的表达。提高Nrf2、Ho-1和Gpx1 mRNA和蛋白的表达。PFD治疗后，肺纤维化小鼠MLF、血清、BALF上清液中col1a1、IL-6水平以及肺组织中ROS、血清和BALF中MDA水平均显著降低，肺炎症细胞浸润及纤维化程度均有所减轻。PFD对IPF的治疗作用涉及调节氧化应激能力的Nrf2/Bach1平衡。该研究为PFD的抗氧化机制提供了新的认识。本文的在线版本（doi:10.1186/s12890-017-0405-7）包含补充材料，可供授权用户使用。
Oxidative stress is one of the important factors involved in the pathogenesis of idiopathic pulmonary fibrosis (IPF). The equilibrium of Nuclear factor-erythroid-related factor 2 (Nrf2)/[BTB (broad-complex, tramtrack and bric-a-brac) and CNC (cap‘n’collar protein) homology 1, Bach1] determines the expression level of antioxidant factors, further regulating the function of oxidation/antioxidation capacity. Pirfenidone (PFD) is one of two currently for IPF therapy approved drugs. PFD regulates intracellular antioxidants, inhibits secretion of inflammatory cytokines and collagen synthesis. However the mechanisms of its antioxidant effects remain elusive. Effects of PFD treatment were studied in mouse lung fibroblasts (MLF) following induction by transforming-growth factor beta 1 (TGF-β1) and in mice following bleomycin-induced lung fibrosis. The mRNA and protein levels of oxidative stress-related factors Nrf2/Bach1 and their downstream antioxidant factors heme oxygenase-1 (Ho-1) and glutathione peroxidase 1 (Gpx1) were determined by RT-PCR and Western blot. Fibrosis-related cytokines interleukin-6 (IL-6) and myofibroblast markers type 1 collagen α1 (COL1A1) levels in supernate of MLF, serum, and bronchoalveolar lavage fluid (BALF) as well as malondialdehyde (MDA) in serum and BALF were detected by ELISA, reactive oxygen species (ROS) generation was measured by 2′,7′- dichlorofluorescin diacetate (DCFH-DA) assay and lung pathological/morphological alterations in mice were observed by HE and Masson to assess the antioxidant mechanism and therapeutic effects on pulmonary fibrosis induced by bleomycin. PFD inhibited Bach1 mRNA and protein expressions in mouse lung fibroblasts induced by TGF-β1 and lung tissues with pulmonary fibrosis induced by bleomycin. Furthermore, it improved Nrf2, Ho-1 and Gpx1 mRNA and protein expressions. After PFD treatment, COL1A1and IL-6 levels in supernate of MLF, serum, and BALF as well as ROS in lung tissues and MDA in serum and BALF from a mouse with pulmonary fibrosis were significantly decreased, and the infiltration of lung inflammatory cells and fibrosis degree were alleviated. Theraputic effects of PFD for IPF were involved in Nrf2/Bach1 equilibrium which regulated the capacity of oxidative stress. The study provided new insights into the antioxidant mechanism of PFD. The online version of this article (doi:10.1186/s12890-017-0405-7) contains supplementary material, which is available to authorized users.