Study On The Effect Of Yiqi Huoxue Xiaozheng Recipe On COPD Fibroblast Proliferation And Inflammatory Factor Expressio

Background:Chronic obstructive pulmonary disease,a common respiratory disease,is a preventable and treatable disease.The prevalence rate is increasing year by year.The patient’s quality of life is low,the mortality rate is high,and the social and economic burden is too heavy.It has become a public health issue of global concern,and its mechanism research has received much attention.Current research shows that the pathogenesis of COPD mainly includes inflammatory reaction,protease/antiprotease imbalance,oxidation/antioxidation imbalance,autoimmunity,alveolar septal cell apoptosis,gene polymorphism,chronic infection of Chlamydia pneumoniae.COPD is characterized by increased levels of inflammatory factors and various proteases.NF-κB can induce a variety of inflammatory cells,such as IL-6,TNF-α,colony-stimulating factor,adhesion molecules and other inflammatory cytokines.Inflammatory mediators participate in many aspects of the chronic inflammatory response of the airway,and promote the development of inflammation to a certain extent,causing pathological changes or tissue damage,and eventually leading to tracheal damage and recurrent inflammation.Even the delay did not heal.Professor Wang Qi proposed the hypothesis of "micro zhengjia obstructs lung collateral",and based on this theory,formulated the treatment of COPD with the method of Yiqi Huoxue Xiaozheng.The previous clinical trials of our group showed that Yiqi Xiaozheng Recipe can improve the lung function and blood rheology index of patients with stable COPD in a certain degree.The previous animal experiments showed that Yiqi Huoxue Xiaozheng Recipe can regulate the airway of rats by COPD model through retrieve the protease/antiprotease imbalance of epithelial cells;Yiqi Huoxue Xiaozheng decoction can also reduce the levels of TNF-α,IL-7 and IFN-γ in serum of rats with COPD and improve the rats’lung function.Based on the previous experimental study,this experiment is to explore the effect of Yiqi Huoxue Xiaozheng Decoction on the proliferation and inflammatory factors of fibroblasts in COPD airway remodeling,and make clear its mechanism of action based on the NF-κB classical signaling pathway from molecular biology.Learn from the perspective of the possible target of Yiqi Huoxue Xiaozheng Decoction.Objective:Experiment 1:Construct a model of LPS stimulation of fibroblast proliferation in vitro.Experiment 2:To explore the essence of the prescription of Yiqi Huoxue Xiaozheng Recipe,based on NF-κB signaling pathway,to explore the possible target of this side in the expression of fibroblast inflammatory factors,for the COPD airway inflammation and airway remodeling which is about the "micro zhengjia obstructs lung collateral" hypothesis.Methods:Experiment 1:MRC-5 cells were intervened in vitro with different concentrations of LPS for 24h to stimulate fibroblast proliferation,and CCK-8 was used to detect cell proliferation.Experiment 2:Yiqi Huoxue Xiaozheng was divided into Yiqi Xiaozheng Tongluo Recipe and Yiqi Huoxue Huatan Recipe.Wistar rats were randomly divided into 4 groups,12 in the blank group and 4 in the Yiqi Huoxuexiao group,4 in Yiqixiaodutongluo group,4 in Yiqi Huoxuehuayu group,corresponding giving medicine to normal saline and Yiqi Huoxue Xiaozheng Recipe,Yiqi Xiaozheng Tongluo Recipe,Yiqi Huoxue Huatan Recipe for 7 days.The blood was collected from the abdominal aorta to prepare the drug-containing serum.The MRC-5 cells were cultured and plated according to the group,LPS,drug-containing serum and budesonide were treated for 24 hours respectively to detect cell proliferation and TGF-β1,TNFα,IL-6,p65,IKKβ expression.Results:Experiment 1:LPS concentration of 100-2000 ng/ml can stimulate the proliferation of fibroblast MRC-5 in vitro,and the best concentration is 500 ng/ml to stimulated cell proliferation.Experiment 2:1.Compared with the model group,Yiqi Huoxue Xiaozheng can inhibit the proliferation of MRC-5 cells stimulated by LPS,inhibit the secretion of TGF-β,TNFα,IL-6 by cells,and inhibit the production of p65 and IKKβ.2.Compared with the model group,Yiqi Xiaozheng Tongluo Recipe can inhibit the proliferation of LPS-stimulated MRC-5 cells,inhibit the secretion of TNF-α and IL-6 by cells,and inhibit the production of p65 and IKKβ.3.Compared with the model group Compared with Yiqi Huoxue Huatan Recipe,it can inhibit the secretion of TNF-α and IL-6 by LPS-stimulated MRC-5 cells and inhibit the production of p65 and IKKβ.4.Compared with the model group,Bud can inhibit LPS-stimulated MRC-5 cell proliferation,inhibiting the secretion of TGF-β,TNF-α,IL-6 by cells,inhibiting the production of p65 and IKKβ.5.Compared with Yiqi Xiaozheng Tongluo group,Yiqi Huoxue Xiaozheng group had more obvious inhibition of MRC-5 cell proliferation and IL-6 expression,for inhibiting the expression of TNF-α,p65 and IKKβ,the two groups were equivalent,Yiqi Xiaozheng Tongluo Recipe has no significant effect on the expression of TGF-β.6.Compared with Yiqi Huoxue Huatan group,Yiqi Huoxue Xiaozheng group had more obvious effects on inhibiting the expression of IL-6,p65 and IKKβ,for inhibiting the expression of TNF-α,the two groups had the same effect,Yiqihuoxuehuayu The group had no significant effect on MRC-5 proliferation and TGF-β expression.7.Compared with the Bud group,the Yiqi Huoxuexiao group had more obvious effects on inhibiting the proliferation of MRC-5 cells and the expression of IL-6,p65 and IKKβ.For inhibiting the expression of TGF-β and TNF-α,the two groups were equivalent.Conclusions:Experiment 1:LPS can stimulate fibroblast proliferation in vitro.Experiment 2:Yiqi Huoxue Xiaozheng can inhibit the proliferation of fibroblasts and the expression of inflammatory factors such as TGF-β,TNF-α and IL-6,which may be related to down-regulating the activity of NF-κB signaling pathway.Budesonide is not as effective as Yiqi Huoxue Xiaozheng in inhibiting the proliferation of fibroblasts and inhibiting the expression of p65,IKKβ and IL-6.As a compound of Yiqi Huoxue Xiaozheng,it inhibits the proliferation and the expression inflammatory factors of fibroblasts More obviously.This experiment confirmed the rationality of the "micro zhengjia of obstructs lung collateral" hypothesis,and confirmed the inhibitory effect of Yiqi Huoxue Xiaozheng Recipe on COPD inflammatory response.