Functions of key genes involved in TGF-β/Smad signaling pathway in progression of pulmonary fibrosis

作者：Huinan YANG, Da Lyu, Le Wang, Chuncheng LIU, Zhiyan JIANG, Hongyu Zhao, Lu Cai · 发表于：DOAJ (DOAJ: Directory of Open Access Journals) · 年份：2022 · DOI：10.11836/jeom21599 · 被引用次数：1 · 研究领域：Interstitial Lung Diseases and Idiopathic Pulmonary Fibrosis

BackgroundAlthough transforming growth factor-β (TGF-β)/Smad signaling pathway is important in regulating the occurrence and development of pulmonary fibrosis, the pathogenesis of pulmonary fibrosis remains elusive. ObjectiveTo explore the functions of genes associated with TGF-β/Smad signaling pathway in the progression of pulmonary fibrosis. MethodsA NIH-3T3 fibroblast model induced by TGF-β1 was established. The experiment samples were divided into a control group and a TGF-β1 treatment group. The control group was exposed to normal saline, while the TGF-β1 treatment group was exposed to 10 ng·mL−1 TGF-β1 for 12 h. The RNAs of the two groups were extracted, sequenced, and analyzed by bioinformatics methods to identify seven key genes in TGF-β pathway, including Dcn, Smad3, Smad7, Fbn1, Thbs1, TGF-β1, and TGF-β3. The gene expression levels of five markers [Collagen1α1, Collagen1α2, α-smooth muscle actin (α-SMA), TGF-β1, and TGF-β3] and the seven key genes were detected by quantitative real-time PCR (qRT-PCR). The proteins of the two groups were extracted. The important marker protein expression levels of Smad3, the phosphorylation of Smad3 (P-Smad3), and α-SMA were detected by Western blotting. At the same time, 30 healthy SPF-grade C57BL/6 mice were randomly divided into three groups, with 10 mice in each group: a control group, a SiO2 inhalation exposure group for 28 d (10 mice), and a SiO2 inhalation exposure group for 56 d (10 mice). The mice in the two treatment groups...