Osteopontin silencing attenuates bleomycin-induced murine pulmonary fibrosis by regulating epithelial-mesenchymal transition
Osteopontin silencing attenuates bleomycin-induced murine pulmonary fibrosis by regulating epithelial-mesenchymal transition
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DOI：
10.1016/j.biopha.2021.111633
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发表时间：
2021-05-08
影响因子：
7.5
通讯作者：
Hirohata, Satoshi
中科院分区：
文献类型：
--
作者：
Hatipoglu, Omer Faruk;Uctepe, Eyyup;Hirohata, Satoshi
特发性肺纤维化（IPF）是间质性肺病最常见和最致命的形式。骨桥蛋白（OPN）是一种具有促炎和促纤维化特性的基质细胞蛋白，在包括IPF在内的几种纤维化疾病中起主要作用; OPN在患者的肺样本中高度上调。在这项研究中，我们使用小干扰RNA（siRNA）敲低博莱霉素（BLM）诱导的肺纤维化（PF）小鼠模型中的OPN，以确定使用OPN siRNA是否是IPF的有效治疗策略。我们发现，在OPN siRNA处理的小鼠标本中，纤维化区域明显较小。在OPN siRNA处理的小鼠中，支气管肺泡灌洗液中肺泡巨噬细胞、中性粒细胞和淋巴细胞的数量也减少。关于上皮-间质转化（EMT）相关蛋白的表达，向BLM处理的小鼠施用OPN-siRNA上调E-cadherin表达并下调波形蛋白表达。此外，在体外，我们孵育人肺泡腺癌细胞系A549与转化生长因子（TGF）-β 1，随后转染细胞与OPN siRNA。我们发现在A549细胞中，TGF-β 1刺激后Col 1A 1、纤连蛋白和波形蛋白显著上调。相反，在TGF-β 1刺激的OPN敲低A549细胞中观察到Col 1A 1、纤连蛋白和波形蛋白mRNA水平下调。因此，在体外和体内，下调OPN有效地减少肺纤维化和EMT变化。总之，我们的结果表明，OPN siRNA对BLM诱导的小鼠PF具有保护作用。我们的研究结果为开发新型IPF靶向治疗策略提供了基础。
Idiopathic pulmonary fibrosis (IPF) is the most common and most deadly form of interstitial lung disease. Osteopontin (OPN), a matricellular protein with proinflammatory and profibrotic properties, plays a major role in several fibrotic diseases, including IPF; OPN is highly upregulated in patients' lung samples. In this study, we knocked down OPN in a bleomycin (BLM)-induced pulmonary fibrosis (PF) mouse model using small interfering RNA (siRNA) to determine whether the use of OPN siRNA is an effective therapeutic strategy for IPF. We found that fibrosing areas were significantly smaller in specimens from OPN siRNA-treated mice. The number of alveolar macrophages, neutrophils, and lymphocytes in bronchoalveolar lavage fluid was also reduced in OPN siRNA-treated mice. Regarding the expression of epithelial-mesenchymal transition (EMT)-related proteins, the administration of OPN-siRNA to BLM-treated mice upregulated E-cadherin expression and downregulated vimentin expression. Moreover, in vitro, we incubated the human alveolar adenocarcinoma cell line A549 with transforming growth factor (TGF)-beta 1 and subsequently transfected the cells with OPN siRNA. We found a significant upregulation of Col1A1, fibronectin, and vimentin after TGF-beta 1 stimulation in A549 cells. In contrast, a downregulation of Col1A1, fibronectin, and vimentin mRNA levels was observed in TGF-beta 1-stimulated OPN knockdown A549 cells. Therefore, the downregulation of OPN effectively reduced pulmonary fibrotic and EMT changes both in vitro and in vivo. Altogether, our results indicate that OPN siRNA exerts a protective effect on BLM-induced PF in mice. Our results provide a basis for the development of novel targeted therapeutic strategies for IPF.