HSG通过Wnt/β-catenin通路抑制COPD大鼠气道成纤维细胞增殖

HSG inhibits the proliferation of fibroblasts in COPD rats by Wnt / β-catenin signaling pathway

目的研究增殖抑制基因(HSG)在大鼠慢性阻塞性肺疾病(COPD)气道重建中与Wnt/β-catenin信号传导通路的关系及其调控气道重建的机制。方法将大鼠随机分为对照组及COPD模型组,模型组采用气道内注射脂多糖加烟熏方法建立。HE染色鉴定COPD组织,测定气道管壁总面积/气道基底膜周长(WAt/Pbm)评价气道重建情况。组织块贴壁法培养大鼠气道成纤维细胞,real time-PCR检测成纤维细胞HSG和β-catenin mRNA表达。Western blot联合ELISA法检测HSG、β-catenin、GSK-3β、TGF-β1和MMP-9蛋白表达。分析HSG、β-catenin基因表达与TGF-β1和MMP-9细胞因子分泌的相关性。结果模型组大鼠小气道重建显著。COPD大鼠气道成纤维细胞中HSG mRNA和蛋白表达均低于对照组。β-catenin mRNA和蛋白表达均高于对照组,P-GSK-3β、TGF-β1和MMP-9蛋白表达也高于对照组。HSG mRNA的表达与β-catenin mRNA表达及与TGF-β1和MMP-9细胞因子分泌呈负相关。结论 HSG可能通过调控Wnt/β-catenin信号通路,抑制大鼠气道成纤维细胞的增殖从而参与气道重建。

Objective Research the relationship between HSG and Wnt / β-catenin signal transduction pathway in airway remodeling in COPD,to elaborate HSG gene regulation mechanism of airway remodeling. Methods Rats were divided into normal control group and COPD model group. COPD model group was intratracheal injection of LPS plus smoking. Used HE staining to identify COPD model. Determination of the total area of the airway wall / airway basement membrane perimeter（ WAt / Pbm） evalution of airway remodeling situation. Cultured the rat airway primary fibroblasts cells in vitro by tissue blocking adhering method. Realtime-PCR were used to detect the HSG andβ-catenin mRNA expression in fibroblasts. Used Western blot assay and ELISA for HSG,β-catenin,GSK-3β,TGF-β1,MMP-9 in small airway fibroblasts. Explored the correlation of HSG,β-catenin with TGF-β1,MMP-9.Results Small airway remodeling in model group was significant. COPD model group airway fibroblasts HSG mRNA,protein expression were lower than the normal group. mRNA,protein expression of β-catenin were higher thannormal group. Expression of P-GSK-3β,TGF-β1 and MMP-9was also higher than that in normal group. HSG mRNA expression and mRNA expression of β-catenin and TGF-β1,MMP-9 secretion of cytokines were negatively correlated. Conclusions The HSG gene may regulate Wnt / β-catenin signaling pathway,whcih resulted in inhibiting the fibroblasts proliferation.