BTB和CNC同源物1抑制通过阻断肺纤维化中的ERK途径改善纤维化和炎症。
BTB and CNC homology 1 inhibition ameliorates fibrosis and inflammation via blocking ERK pathway in pulmonary fibrosis.
作者信息
Liu Yuan, Wang Yongfu, Lu Fuai, Wang Le, Miao Liu, Wang Xiaoyuan
机构信息
Department of Rheumatology, Liuzhou People's Hospital , Liuzhou , China.
Department of Rheumatology, First Affiliated Hospital of Baotou Medical College , Baotou , China.
出版信息
Exp Lung Res. 2021 Feb-Mar;47(2):67-77. doi: 10.1080/01902148.2020.1849448. Epub 2020 Nov 26.
OBJECTIVE
Patients with idiopathic pulmonary fibrosis (IPF) are still suffering from unfavorable survival. BTB and CNC homology1 (Bach1) is a regulator of oxidative stress and participates in the pathogenesis of multiple lung diseases. Thus, this study aimed to determine the effect of Bach1 knockdown on fibrosis and inflammation in pulmonary fibrosis (PF) mice and cell models.
METHODS
Bleomycin induced PF mice were constructed and treated with Bach1 siRNA adenovirus (BLM + Bach1 siRNA group), control siRNA adenovirus (BLM + Control siRNA group) or normal saline (BLM group), then lung tissues were collected for Bach1 expression detection, H&E staining and Masson's trichrome staining. Afterwards, collagen type I alpha 1 chain (COL1A1) and interleukin-6 (IL-6) expressions in serum and bronchoalveolar lavage fluid (BALF) were examined. Subsequently, mouse lung fibroblasts (MLFs) were collected from PF mice and treated with TGF-β1 to construct PF cell model, which was treated with Bach1 siRNA adenovirus (TGF-β1 + Bach1 siRNA group) and MAP kinase (ERK) inhibitor U0126 alone (TGF-β1 + U0126 group) or in combination (TGF-β1 + U0126 + Bach1 siRNA group), then alpha-smooth muscle actin (α-SMA), fibronectin 1 (Fn1), COL1A1, IL-6 expressions and cell viability were detected.
RESULTS
Lung tissue Bach1 mRNA and protein expressions were upregulated in PF mice compared to control mice. Bach1 knockdown reduced lung fibrosis (displayed by Masson's trichrome staining) and inflammation (displayed by H&E staining), then downregulated serum and BALF expressions of COL1A1 and IL-6 in PF mice. Subsequently, in PF cell model, Bach1 knockdown blocked ERK pathway, but did not affect Smads, c-Jun N-terminal kinase (JNK) or thymoma viral proto-oncogene 1 (Akt) pathways. Further experiments revealed that Bach1 knockdown repressed cell viability, α-SMA, Fn1, IL-6 and COL1A1 expressions in PF cell model, then ERK inhibition by U0126 enhanced these effects.
CONCLUSIONS
Bach1 is involved in the PF pathogenesis via modulating ERK signaling pathway.
目的
特发性肺纤维化（IPF）患者的生存状况仍然不佳。BTB和CNC同源蛋白1（Bach1）是氧化应激的调节因子，参与多种肺部疾病的发病机制。因此，本研究旨在确定敲低Bach1对肺纤维化（PF）小鼠和细胞模型中纤维化和炎症的影响。
方法
构建博来霉素诱导的PF小鼠模型，并分别用Bach1 siRNA腺病毒（博来霉素+Bach1 siRNA组）、对照siRNA腺病毒（博来霉素+对照siRNA组）或生理盐水（博来霉素组）进行处理，然后收集肺组织进行Bach1表达检测、苏木精-伊红（H&E）染色和Masson三色染色。之后，检测血清和支气管肺泡灌洗液（BALF）中Ⅰ型胶原α1链（COL1A1）和白细胞介素-6（IL-6）的表达。随后，从PF小鼠中分离出小鼠肺成纤维细胞（MLF），并用转化生长因子-β1（TGF-β1）处理构建PF细胞模型，该模型分别用Bach1 siRNA腺病毒（TGF-β1+Bach1 siRNA组）、丝裂原活化蛋白激酶（ERK）抑制剂U0126单独处理（TGF-β1+U0126组）或联合处理（TGF-β1+U0126+Bach1 siRNA组），然后检测α平滑肌肌动蛋白（α-SMA）、纤连蛋白1（Fn1）、COL1A1、IL-6的表达及细胞活力。
结果
与对照小鼠相比，PF小鼠肺组织中Bach1 mRNA和蛋白表达上调。敲低Bach1可减轻肺纤维化（通过Masson三色染色显示）和炎症（通过H&E染色显示），并下调PF小鼠血清和BALF中COL1A1和IL-6的表达。随后，在PF细胞模型中，敲低Bach1可阻断ERK通路，但不影响Smads、c-Jun氨基末端激酶（JNK）或胸腺瘤病毒原癌基因1（Akt）通路。进一步实验表明，敲低Bach1可抑制PF细胞模型中的细胞活力、α-SMA、Fn1、IL-6和COL1A1的表达，而U0126抑制ERK可增强这些作用。
结论
Bach1通过调节ERK信号通路参与PF的发病机制。
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