细胞增殖抑制基因调控慢性阻塞性肺疾病大鼠Wnt/PCP通路抑制气道成纤维细胞增殖

Hyperplasia suppressor gene inhibits proliferation of airway fibroblasts by regulating Wnt/PCP signaling pathway in COPD rats

目的探讨细胞增殖抑制基因(HSG)在大鼠慢性阻塞性肺疾病(COPD)气道重建中Wnt/PCP通路的作用及其机制。方法将大鼠分为对照组和COPD模型组;采用烟熏+气道内注射脂多糖建立;取气管做小气道成纤维细胞原代培养,第3代细胞系用于实验。HE染色鉴定肺组织是否造模成功。ELISA检测培养上清液中MMP-9、PDGF和TGF-β1的表达;real-time PCR检测成纤维细胞中HSG、PDGF、TGF-β1和RhoA的mRNA的表达;Western blot法检测成纤维细胞HSG和RhoA蛋白表达。结果模型组成纤维细胞上清液中PDGF、TGF-β1和MMP-9表达显著高于对照组(P<0.01);模型组成纤维细胞中HSG mRNA及蛋白表达显著低于对照组(P<0.01);RhoA蛋白及PDGF、TGF-β1和RhoA mRNA表达显著高于对照组(P<0.01)。HSG表达与MMP-9、PDGF、TGF-β1和RhoA表达呈负相关(P<0.01)。结论HSG可能通过调控Wnt/PCP通路,抑制大鼠气道成纤维细胞的增殖从而参与气道重建。

Objective To research the relationship between HSG and Wnt/PCP signal transduction pathway in airway remodeling in COPD,and to elaborate HSG gene regulation mechanism of airway remodeling. Methods Rats were divided into COPD model group and normal control group, Small airway fibroblasts were taken to perform primary cell culture, the third generation of cell lines was used for experiments. Used HE staining to identify COPD model, ELISA was used to detect the expression of MMP-9, PDGF and TGF-β1 in small airways;real-time PCR was used to detect the mRNA expression of the HSG, PDGF, TGF-β1, Western blot was used to detect the expression of the HSG, RhoA protein in fibroblast;RhoA protein in fibroblast. Results Supernatant PDGF, TGF-β1 and MMP-9 in model group airway fibroblasts were higher than normal control group(P<0.01);HSG protein and mRNA in model group airway fibroblasts were lower than normal control group(P<0.01);but RhoA protein and the expression of PDGF, TGF-β1, RhoA gene mRNA were higher than normal control group(P<0.01);The expression of HSG was negatively correlated with that of MMP-9, PDGF, TGF-β1 and RhoA(P<0.01). Conclusions The HSG gene may regulate Wnt/PCP signaling pathway,which in turn inhibitats the fibroblasts proliferation.