Effect Of XueFuZhuYu Decoction On TLR4/MyD88/NF-κB Signaling Pathway In Chronic Obstructive Pulmonary Disease

Objective:Chronic obstructive pulmonary disease(COPD)is a common pulmonary disease in clinic,with the main pathological features of chronic inflammation invading the airways,pulmonary parenchyma and pulmonary vessels.TLR4/MyD88/NF-κB signaling pathway is particularly important for the development of COPD by promoting the release of airway inflammatory factors and increasing mucin secretion.COPD belongs to the category of "lung distension" in traditional Chinese medicine.XueFuZhuYu decoction has achieved great efficacy in the treatment of COPD and inhibited the release of airway inflammatory factors.However,its mechanism is still unclear.There are few study about the therapeutic role in airway inflammation reaction and its related pathways.In This experiment,mice and human peripheral venous blood is the object of study.We try to explore the Intervention effect of XueFuZhuYu decoction on TLR4/MyD88/NF-κB signaling pathway and its possible mechanisms,provide more theoretical basis for clinical evaluation and treatment of COPD.Methods:The first part is to explore the role of XueFuZhuYu decoction at release inflammatory cytokines in mice with COPD and the lung tissue of TLR4、MyD88、NF-κB protein expression.Cigarette smoke joint intranasal LPS method for COPD mice model,last for 120 days.After the success of the model building,the COPD mice are divide into six group:blank group,model group,positive drug group,low dose group of XueFuZhuYu decoction,medium dose group of XueFuZhuYu decoction and high dose group of XueFuZhuYu decoction.After 30 days of drug delivery,the daily activity,diet,respiratory rate,secretion,hair color and body weight of mice in each group were observed.HE staining was used to observe the changes of trachea and bronchial structure,alveolar dilatation,deformation and fusion,infiltration of neutrophils in lung parenchyma,vascular deformation and smooth muscle hyperplasia in each group.ELISA is used to detect the levels of TNF-αIL-1β,IL-6,IL-8 and IL-17 in mice BALF and serum.The levels of TLR4,MyD88 and NF-κB in mice lung tissues are detected by immunohistochemistry.The second part is to explore the function of XueFuZhuYu decoction in regulating human peripheral venous blood mononuclear cells(PBMCs)TLR4/MyD88/NF-κB signaling pathways.10ml peripheral venous blood of 15 cases of patients with COPD and 15 cases of ordinary people are included in the study.We using density gradient centrifugation separation get the PBMCs and determine PHA and therapeutic drug concentration.The specimen are divide into six group:the blank+PHA group,blank+ PHA+XueFuZhuYu decoction group,COPD+PHA group,COPD+PHA+XueFuZhuYu decoction group.Cell culture is conducted.ELISA is used to detect the levels of TNF-αIL-1β,IL-6,IL-8 and IL-17 in PBMCs.The mRNA and protein expressions of TLR4,MyD88 and NF-κB in PBMCs cells are detected by RT-PCR and western blot.Conclusions:Part 1:XueFuZhuYu decoction can improve the general situation of COPD mice,prevent the mice from breathing too fast,reduce cough,reduce secretions,and avoid excessive weight loss.HE staining under electron microscopy showed that XueFuZhuYu decoction could reduce the pathological changes of small airways,alveoli and blood vessels in mice,and reduce the accienulation of inflammatory cells.Moreover,xuefuyu decoction could reduce the expression of TNF-αIL-1β,IL-6,IL-8 and IL-17 in BALF and serum of COPD mice,with statistically significant differences(P<0.05).Immunohistochemistry observed that XueFuZhuYu decoction could reduce the expression of TLR4,MyD88 and NF-κB proteins in lung tissues of COPD micePart 2:XueFuZhuYu decoction can reduce the level of TNF-αIL-1β,IL-8 and IL-17 in the peripheral venous blood PBMCs,with statistically significant differences(P<0.05).A slight decrease in IL-6 without statistically significant differences(P>0.05).RT-PCR and western blot results showed that XueFuZhuYu decoction could reduce the expression of TLR4,MyD88 and NF-κB proteins in PBMCs cells,and the difference was statistically significant(P<0.05).Results:XueFuZhuYu decoction can reduce the levels of inflammatory cytokines TNF-αIL-1β,IL-8 and IL-17in mice BALF and serum,and human peripheral blood PBMCs.The mechanism may be is the control of the TLR4/MyD88/NF-κB signaling pathway,which plays a therapeutic role in chronic obstructive pulmonary disease.