Department of Respiratory Medicine, The Second Affiliated Hospital of Guizhou College of Traditional Chinese Medicine, Guiyang, Guizhou 550003, P.R. China.
Department of Research and Teaching, The Second Affiliated Hospital of Guizhou College of Traditional Chinese Medicine, Guiyang, Guizhou 550003, P.R. China.
Mol Med Rep. 2019 Sep;20(3):2754-2762. doi: 10.3892/mmr.2019.10504. Epub 2019 Jul 15.
The present study aimed to investigate the effects of hyperplasia suppressor gene (HSG) overexpression on the activation of airway fibroblasts in a rat model of chronic obstructive pulmonary disease (COPD) and assess the underlying molecular mechanisms. The rat model of COPD was established by injection of papain and confirmed by hematoxylin and eosin staining. Airway fibroblasts were identified using immunofluorescence, and HSG expression was facilitated by an HSG vector. Cell viability, apoptosis and the levels of matrix metallopeptidase‑9 (MMP‑9), platelet‑derived growth factor (PDGF), and transforming growth factor‑β1 (TGF‑β1) were measured via Cell Counting Kit‑8, flow cytometry and ELISA analyses, respectively, and potential mechanisms were detected by reverse transcription‑quantitative polymerase chain reaction and western blotting. Airway fibroblasts from COPD rats were isolated and identified based on vimentin expression. Compared with the control group, HSG overexpression reduced cell viability, promoted apoptosis, and reduced the protein levels of TGF‑β1, MMP‑9 and PDGF. Additionally, HSG overexpression reduced β‑catenin and Ras homology family member A (RhoA) expression at both the mRNA and protein levels. Conversely, Wnt signaling pathway agonists lithium chloride (LiCl) and 4‑ethyl‑5,6‑dihydro‑5‑methyl‑(1,3)dioxolo(4,5‑j)phenanthridine (HLY78), significantly reduced the effects of HSG overexpression (P<0.05 vs. HSG). Cell viability in the HSG + LiCl and HSG + HLY78 groups was increased, whereas apoptosis was reduced compared with HSG treatment alone. The protein levels of TGF‑β1, MMP‑9 and PDGF were also decreased in the HSG + LiCl and HSG + HLY78 groups compared with the HSG group (P<0.05). Furthermore, the expression of β‑catenin and RhoA was higher in the HSG + LiCl and HSG + HLY78 groups compared with the HSG group (P<0.05). Collectively, the results indicated that HSG overexpression inactivated airway fibroblasts from COPD by inhibiting the Wnt signaling pathway.
本研究旨在探讨增生抑制基因（HSG）过表达对慢性阻塞性肺疾病（COPD）大鼠模型气道成纤维细胞激活的影响，并评估其潜在的分子机制。采用木瓜蛋白酶注射法建立 COPD 大鼠模型，并通过苏木精-伊红染色进行验证。采用免疫荧光法鉴定气道成纤维细胞，通过 HSG 载体促进 HSG 表达。通过细胞计数试剂盒-8、流式细胞术和酶联免疫吸附试验分别检测细胞活力、凋亡以及基质金属蛋白酶-9（MMP-9）、血小板衍生生长因子（PDGF）和转化生长因子-β1（TGF-β1）水平，采用逆转录-定量聚合酶链反应和蛋白质印迹法检测潜在机制。基于波形蛋白表达从 COPD 大鼠中分离和鉴定气道成纤维细胞。与对照组相比，HSG 过表达降低了细胞活力，促进了细胞凋亡，并降低了 TGF-β1、MMP-9 和 PDGF 的蛋白水平。此外，HSG 过表达降低了 β-连环蛋白和 Ras 同源家族成员 A（RhoA）在 mRNA 和蛋白水平的表达。相反，Wnt 信号通路激动剂氯化锂（LiCl）和 4-乙基-5,6-二氢-5-甲基-(1,3)二氧杂环(4,5-j)菲啶（HLY78）显著减轻了 HSG 过表达的影响（与 HSG 组相比，P<0.05）。与单独 HSG 处理相比，HSG + LiCl 和 HSG + HLY78 组的细胞活力增加，而凋亡减少。与 HSG 组相比，HSG + LiCl 和 HSG + HLY78 组 TGF-β1、MMP-9 和 PDGF 的蛋白水平也降低（P<0.05）。此外，与 HSG 组相比，HSG + LiCl 和 HSG + HLY78 组的 β-连环蛋白和 RhoA 表达更高（P<0.05）。综上所述，研究结果表明，HSG 过表达通过抑制 Wnt 信号通路使 COPD 气道成纤维细胞失活。
