Yu-Ping-Feng-San, Si-Wu-Tang And The Combined Formula Interfering COPD Airway Remodeling In Preliminary Research On The Molecular Mechanism

ObjectiveBased on Yunnan province famous traditional Chinese medicine in the treatment of chronic obstructive pulmonary disease（COPD） ancient prescription Yu-Ping-Feng-San（YPFS） and Si-WuTang（SWT） are effective in clinical, this study will be on the prescription of action mechanism are discusse from animal models and somatic cells, we select indicators of COPD related TGF-β₁/Smad2,3, proinflammatory factor and collagen expression, to study the prescription intervention molecular action mechanisms of COPD airway remodeling.Methods1.Establish a rat model of COPD, after the intervention of YPFS, SWT and the combined formula to detect the phosphorylation level of Smad2,3 in the TGF-β₁/Smad2,3 signaling pathways,proinflammatory factor IL-6,IL-1β, and collagen COL3A1 expression.2.Establish the bronchial epithelial cells（Beas-2b） inflammation model, after the intervention of YPFS, SWT and the combined formula to detect the phosphorylation level of Smad2,3 in the TGF-β₁/Smad2,3 signaling pathways, proinflammatory factor IL-6, collagen COL3A1 expression.3.In the bronchial epithelial cells（Beas-2b） inflammation model, by RNAi to silence smad2 gene in Beas-2b cells, to explore whether YPFS, SWT and the combined formula adjust TGF-β₁/Smad2,3 signaling pathways play a role of anti-inflammatory and cut the expression of collagen COL3A1.Results1.The function of YPFS, SWT and the combined formula in COPD in rats model of is as follows:1.1 Pathological results show that inflammatory cell infiltrate, the alveolar was damaged, and the septal of alveolar is thickening in rat’s lung tissue in model group. After the intervention of YPFS, SWT and the combined formula, infiltration of inflammatory cells is less than the model group in lung tissue of rats.1.2 The phosphorylation level of Smad2,3 in the TGF-β₁/Smad2,3 signaling pathways show that YPFS, SWT and the combined formula group phosphorylation level of Smad2,3 is obviously lower than the model group in the lung tissue（P<0.01）. The effect of combined formula is better than single prescription（P<0.01）.1.3 The expression of proinflammatory factor IL-6,IL-1β show that the expression of IL-6 is obviously lower in YPFS, SWT and combined formula group than the model group in the lung tissue（P<0.01）. The mRNA level of proinflammatory factor IL-6,IL-1β mRNA is obviously lower in YPFS group than the model group in the lung tissue（P<0.05）.1.4 The expression of collagen COL3A1 show that the expression of collagen COL3A1 is obviously lower in YPFS, SWT and the combined formula group than the model groupin in the lung tissue（P<0.01）. The effect of combined formula is better than single prescription（P<0.01）.2. The function of YPFS, SWT and the combined formula in cell inflammation model of is as follows:2.1 Under the light microscopy show that Beas-2b cells was stimulated by lipopolysaccharide or nicotine,which made the number of apoptotic cells was increased. After the intervention of YPFS, SWT and the combined formula, cell apoptosis is inhibited.2.2 The phosphorylation level of Smad2,3 in the TGF-β₁/Smad2,3 signaling pathways show that YPFS, SWT and the combined formula group phosphorylation level of Smad2,3 is obviously lower than the model group in the cells（P<0.01）. The effect of combined formula is better than single prescription（P<0.01）.2.3 The expression of proinflammatory factor IL-6 show that the expression of IL-6 is obviously lower in YPFS,SWT and combined formula group than the model group in the cells（P<0.01）.2.4 The expression of collagen COL3A1 show that the expression of collagen COL3A1 is obviously lower in YPFS, SWT and the combined formula group than the model groupin in the cells（P<0.01）.3. After administration RNAi the smad2 gene in Beas-2b cells, after the intervention of YPFS,SWT and the combined formula, the expression of proinflammatory factor IL-6 and collagen COL3A1 is lower in YPFS, SWT and combined formula group than the model group in the cells,but reduce the amplitude is smaller than no administration RNAi the smad2 gene.Conclusion1. YPFS and SWT inhibit the level of phosphorylation of Smad2,3 in different degrees,the expression of proinflammatory factor IL-6,IL-1β and collagen COL3A1 are decreased, to relieve the process of airway remodeling and inflammation.2. The effect is more obvioue in combined formula than YPFS or SWT single formula on inhibition of Smad2,3-phosphorylation.3. After administration RNAi the smad2 gene in Beas-2b cells, the TGF-β₁/Smad2,3 signaling pathway was crippled, the anti-inflammatory effect of YPFS, SWT and the combined formula was decreased and collagen deposition was increased. It shows that YPFS, SWT and the combined formula regulates the TGF-β₁/Smad2,3 signaling pathway to lower the inflammation and reduce the collagen expression.