Wong Steven, Botelho Fernando M, Rodrigues Rebecca M, Richards Carl D
McMaster Immunology Research Centre, Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada.
Lab Invest. 2014 Sep;94(9):1003-16. doi: 10.1038/labinvest.2014.81. Epub 2014 Jun 16.
Adverse health outcomes in pulmonary fibrosis are associated with extracellular matrix (ECM) accumulation. Although transforming growth factor-β (TGF-β) has been reported to be an important regulator of fibrosis pathogenesis, TGF-β-independent pathways may also be involved. Here, we investigated responses of putative relatively fibrosis-resistant BALB/c mice to transient pulmonary overexpression of oncostatin M (OSM) using an adenovirus vector encoding OSM (AdOSM) and compared responses with the relatively fibrosis-prone C57Bl/6 strain. Interestingly, BALB/c mice showed similar ECM accumulation and collagen 1A1 and 3A1 mRNA elevation to C57Bl/6 mice 7 days after endotracheal administration of AdOSM. TGF-β1 mRNA levels and pSMAD2 signal were not regulated in either strain in total lung extracts. In contrast to C57Bl/6 mice, BALB/c mice lacked eosinophil, Th2 cytokine, and pro-inflammatory cytokine elevation in the broncholveolar space. OSM overexpression induced STAT3 activation and SMAD1/5/8 signaling suppression in lung from both mice strains, which was associated with a downregulation of BMPR2 and BMP ligands, and increased expression of the BMP antagonist gremlin. Although we also observed STAT3 activation and SMAD1/5/8 signaling suppression in mouse lung fibroblast cultures in vitro upon OSM stimulation, immunohistochemistry analyses indicated that the AdOSM-induced pSMAD1/5/8 signal suppression was primarily localized to the airway epithelium. Other gp130 cytokines including IL-6, LIF, CT-1, but not IL-31, also induced STAT3 activation and SMAD1/5/8 signaling suppression in C10 mouse lung epithelial cells and BEAS 2B bronchial epithelial cells, and we found that pharmacological inhibition of STAT3 activation reversed OSM-induced SMAD1/5/8 signaling suppression in vitro. The results demonstrate that OSM induces ECM accumulation in fibrosis-resistant BALB/c mouse lung in the absence of Th2 inflammation or TGF-β signaling, and highlight a dichotomy of STAT3 activation versus SMAD1 suppression in this process.
肺纤维化中的不良健康后果与细胞外基质(ECM)积累有关。尽管转化生长因子-β(TGF-β)已被报道为纤维化发病机制的重要调节因子,但TGF-β非依赖性途径可能也参与其中。在此,我们使用编码制瘤素M(OSM)的腺病毒载体(AdOSM)研究了假定相对抗纤维化的BALB/c小鼠对短暂性肺内OSM过表达的反应,并将其反应与相对易发生纤维化的C57Bl/6品系进行比较。有趣的是,在气管内给予AdOSM 7天后,BALB/c小鼠与C57Bl/6小鼠表现出相似的ECM积累以及胶原蛋白1A1和3A1 mRNA升高。在全肺提取物中,两种品系的TGF-β1 mRNA水平和pSMAD2信号均未受到调节。与C57Bl/6小鼠不同,BALB/c小鼠在支气管肺泡腔中缺乏嗜酸性粒细胞、Th2细胞因子和促炎细胞因子的升高。OSM过表达在两种小鼠品系的肺中均诱导STAT3激活和SMAD1/5/8信号抑制,这与BMPR2和BMP配体的下调以及BMP拮抗剂gremlin的表达增加有关。尽管我们在体外OSM刺激的小鼠肺成纤维细胞培养物中也观察到STAT3激活和SMAD1/5/8信号抑制,但免疫组织化学分析表明,AdOSM诱导的pSMAD1/5/8信号抑制主要定位于气道上皮。包括IL-6、LIF、CT-1但不包括IL-31在内的其他gp130细胞因子,也在C10小鼠肺上皮细胞和BEAS 2B支气管上皮细胞中诱导STAT3激活和SMAD1/5/8信号抑制,并且我们发现STAT3激活的药理学抑制在体外逆转了OSM诱导的SMAD1/5/8信号抑制。结果表明,在不存在Th2炎症或TGF-β信号的情况下,OSM诱导抗纤维化的BALB/c小鼠肺中ECM积累,并突出了在此过程中STAT3激活与SMAD1抑制的二分法。