苗药紫金牛、铁扫帚组方干预慢性阻塞性肺疾病大鼠气道Wnt通路RhoA基因与TGF-β_(1)、MMP-9的相关性研究

Effect of Purple Bull and Iron Broom on the expression of RhoA gene in airway Wnt pathway and the expression of TGF-β_(1) and MMP-9 in rats with chronic obstructive pulmonary disease

目的研究苗药紫金牛、铁扫帚组方干预COPD大鼠气道Wnt通路RhoA基因与TGF-β_(1)、MMP-9的相关性,从而探讨苗药紫金牛、铁扫帚配方干预COPD大鼠气道重塑的调控机制。方法构建COPD大鼠模型,进行气道成纤维细胞的培养,采用ELISA检测气道成纤维细胞上清液中TGF-β_(1)、MMP-9细胞因子的含量,并运用Westernblot方法检测气道成纤维细胞中RhoA基因蛋白的表达,从而研究RhoA基因表达与TGF-β_(1)、MMP-9细胞因子的相关性及苗药紫金牛、铁扫帚的影响。结果与正常组相比,COPD大鼠模型组气道成纤维细胞上清液中TGF-β_(1)、MMP-9的含量升高,同时RhoA基因在也是表达增高。经苗药紫金牛、铁扫帚治疗后,细胞因子TGF-β_(1)、MMP-9与RhoA基因的表达则降低,从而得出TGF-β_(1)、MMP-9细胞因子与RhoA基因呈正相关。结论COPD大鼠气道成纤维细胞上清液中TGF-β_(1)、MMP-9细胞因子是COPD大鼠气道重建中成纤维细胞增殖的关键因子。而RhoA基因对组织纤维化有关,且能调控细胞因子TGF-β_(1)、MMP-9。苗药紫金牛、铁扫帚能够抑制炎性因子TGF-β_(1)、MMP-9的表达及WNT通路的RhoA基因的增殖,从而寻求苗药紫金牛、铁扫帚配方干预COPD气道重塑的机制。

Objective To examine the expression of RhoA gene in small airway fibroblasts of chronic obstructive pulmonary disease(COPD)rats before and after the intervention of Purple Bull and Iron Broom,and explore the regulatory mechanism of Chinese herbal medicine Purple Bull and iron broom formula on airway remodeling in(COPD)rats.Methods The content of TGF-β_(1) and MMP-9 in the supernatant of airway fibroblasts was detected by enzyme linked immunoassays(ELISA)and western blot(WB)was used to detect the expression of RhoA gene pro-tein in airway fibroblasts,so as to study the correlation between RhoA gene expression and TGF-β_(1),MMP-9 cyto-kines and the effect of Purple Bull and iron broom.Results Compared with the normal group,the contents of TGF-p1 and MMP-9 in the supernatant of airway fibroblasts in the COPD model group were increased,and the expression of RhoA gene was also increased.After treatment with Purple Bull and iron broom,the expression of cytokines TGF-β_(1),MMP-9 and RhoA gene were decreased,which indicated that TGF-β_(1),MMP-9 were positively correlated with RhoA gene.Conclusion TGF-pl and MMP-9 cytokines in the supernatant of airway fibroblasts from COPD rats are the key factors in the proliferation of airway fibroblasts in COPD rats.RhoA gene is involved in tissue fibrosis,and can regulate cytokines such as TGF-β_(1) and MMP-9.Purple Bull and iron broom could inhibit the expression of TGF-β_(1),MMP-9 and the proliferation of RhoA gene in Wnt pathway,so as to explore the mechanism of the formula of Purple Bull and iron broom in the intervention of COPD airway remodeling.