COPD模型大鼠TGF-β1/Smads通路炎性蛋白的改变

The change of inflammatory proteins in TGF-β1/Smads signaling pathway in COPD model rats

目的观察烟雾暴露(CS)联合脂多糖(LPS)气管内滴注方法致慢性阻塞性肺病(COPD)模型大鼠TGF-β1/Smad通路中相关炎性蛋白的表达情况。方法将20只鼠龄8周的SPF级雄性Wistar大鼠随机分为对照组和模型组,每组各10只。模型组大鼠采用CS+LPS方法建立COPD模型。采用HE染色法观察肺组织形态改变,酶联免疫吸附(ELISA)法检测外周血血清白细胞介素-17(IL-17)、白细胞介素-10(IL-10)及转化生长因子-β1(TGF-β1)含量,Western blot法测定肺组织TGF-β1、Smad2/3及Smad7磷酸化水平,RT-PCR法测定肺组织中TGF-β1、Smad2/3及Smad7mRNA表达。结果与对照组比较,模型组大鼠支气管及肺泡组织结构紊乱,部分结构消失;肺组织中Smad2/3mRNA表达明显降低(P <0.01);Smad7、IL-10及TGF-β1mRNA表达量明显升高(P<0.01),p-Smad2/3磷酸化明显降低(P <0.01);p-Smad7及TGF-β1磷酸化明显升高(P <0.01);模型组大鼠外周血IL-17水平明显下降,IL-10及TGF-β1水平明显升高(P <0.01),结论 COPD模型大鼠肺组织中TGF-β1/Smads通路处于激活状态,为气道炎症性疾病,特别是COPD的防治提供新的思路和方向。

Objective To observe the expression of related inflammatory proteins in the TGF-β1/Smad pathway in the rats with chronic obstructive pulmonary disease(COPD)induced by cigarette smoking(CS)and tracheal driping lipopolysaccharide(LPS).Methods Twenty 8 week-old male SPF Wistar rats were divided into 2 groups randomly:Blank group and Model group with 10 rats in each group.CS+LPS method was used to establish the COPD model.The method of Hematoxylin-Eosin(HE)staining was used to observe the morphological changes in lung tissue,ELISA method was to detect the serum IL-17,IL-10 and TGF-β1 in peripheral blood,Western Blot method was to determinate expression of p-TGF-β1,p-Smad2/3 and p-Smad7 in lung tissue,RT-PCR method was to determinate TGF-β1,Smad2/3 and Smad7 mRNA expression in lung tissue.Results Compared with the Blank group,the bronchial and alveolar tissues were disordered and some structures disappeared in the Model group.In the Model group,the expression of Smad2/3 mRNA was decreased(P<0.01);the expression levels of Smad7,IL-10 and TGFβ-1 mRNA were increased(P<0.01);the phosphorylation of Smad2/3 decreased(P<0.01);the phosphorylation of Smad7 and TGF-β1 increased(P<0.01);the levels of IL-17 decreased,and IL-10 and TGF-β1 increased(P<0.01).Conclusion The activation of TGF-β1/Smads pathway is confirmed in COPD model rat,which provides a new idea and direction for the prevention and treatment of airway inflammatory diseases,especially COPD.