The Impact Of Systemic Inflammatory Response Airway Remodeling, Pulmonary Vascular Reconstruction In Copd

Objective:To detect the changes of inflammatory cytokines expression in the development of COPD and To assess the degree of systemic inflammatory response in COPD,Find out the relationship between systemic inflammatory response and airway remodeling, also the relationship between systemic inflammatory response and the pulmonary vascular remodeling. To understand the effect of PDTC to inflammatory cytokines expression in animal mode lung tissue of COPD. To research the changes of airway remodeling and pulmonary vascular remodeling after use PDTC.Find out the effect of systemic inflammatory response in the development of COPD and the influence of the PDTC to airway remodeling and pulmonary vascular remodeling in COPD. Giving a new way to the treatment of COPD.Methods:There are two part of this research. Part1:To detect the changes of inflammatory cytokines(high mobility group protein B1,the nuclear factor-Kb, Tumor necrosis facto-a,Interleukin-8, Vascular endothelial growth factor)expression in the development of COPD. Find out the relationship between systemic inflammatory response and the airway and pulmonary vascular remodeling. The experiment uses the smog exposition with the low oxygen to make the animal mode of COPD.(There are three in this part of the research,8SD rat in each group randomly):Healthy control group (group A):0.2ml saline water was installed intratracheally once at first and forteenth day for all rats, testing after6weeks. COPD group (group B):200μg LPS was installed intratracheally once at first and forteenth day and the rats were exposed to cigarette smoke lh/d for6weeks. low oxygen with COPD group (group C):200μg LPS was installed intratracheally once at first and forteenth day and the rats were exposed to cigarette smoke1h/d for6weeks. In last2weeks chronic hypoxia (FiO2=0.18)8h/d were performed simultaneously.Part2:To understand the effect of NF-κB inhibitor pyrrolidine two dithiocarbamate (PDTC)), to inflammatory cytokines (HMGB1, IL-8, TNF-a, VEGF, NF-κB) expression in animal mode of COPD. To research the changes of airway and pulmonary vascular remodeling after use PDTC.(There are three group in this part of the research,8SD rat in each group randomly):PDTC intervention group include:A1(Healthy control group),B1(COPD intervented group), C1(low oxygen with COPD intervented group). Animal model were made the same as the corresponding group, PDTC100mg-kg-1·d-1were injected into the rats in each drug group from the third week. The saline water which was the same dose of PDTC was injected into the rats in A1group from the third week.We examine the following target of rat in each group after animal model made completely:HE staining of lung tissue and VG+Victoria blue triple staining was used to observe the airway and pulmonary vascular remodeling in pathological changes, includeing MT%and MA%,WT%and WA%. Using different methods to detect the expression of various inflammatory factors: Western blot detect the NF-κB protein expression. RT-PCR and Western blot were used to detect HMGB1mRNA and protein expression in each group in lung tissue, ELISA was used to detect TNF-a, IL-8, VEGF expression in lavage fluid (BALF) and serum in each group. Analysis of the expression of various inflammatory factors and airway remodeling, pulmonary vascular remodeling related indicators, as well as expression of PDTC on various inflammatory factors.Results:1.The target of airway remodeling(MT%and MA%):Compare to group A, the target of airway remodeling of group B and C were higher than that of group A(P<0.05). After using PDTC, the target of airway remodeling of group B1and C1were significantly decreased than that of the corresponding group B and C(P<0.05). The above target of group Al which were used saline were no different to that of the corresponding group A(P>0.05).2. The target of pulmonary vascular remodeling(WT%and WA%): Compare to group A, Pulmonary vascular remodeling expressed as the percentage of muscular arteries were increase in group B, But muscle type of artery wall thickness has not been increased.The target of pulmonary vascular remodeling of group C were higher than that of group A(P<0.05). After using PDTC, the target of pulmonary vascular remodeling of group B1and C1were significantly decreased than that of the corresponding group B and C(P<0.05). The above target of group A1which were used saline were no different to that of the corresponding group A(P>0.05).3. The expression of inflammatory factors:3.1The expression of NF-κB Western blot showed that NF-κB protein expression of group B and C were higher than that of group A(P<0.05); After using PDTC NF-κB protein expression of group B1and C1were significantly decreased than that of the corresponding group B and C(P<0.05).3.2The expression of HMGB1RT-PCR and Western blot shows that in group B, C, of HMGB1mRNA, protein expression compared with group A were significantly increased(P<0.05), The expression of NF-κB protein in group C is higher than group B(P<0.05). After using PDTC HMGB1mRNA and protein expression of group B1and C1were significantly decreased than that of the corresponding group B and C (P<0.05).3.3The expression of TNF-a, IL-8, VEGF ELISA results showed that These above target include concentration of TNF-a,IL-8, VEGF in BALF and in serum of group B and C were all higher than that of group A ((P<0.05). After using PDTC, Compared to the group B and C,the expression of TNF-a, IL-8, VEGF in BALF and in serum of group B1and C1, were decreased(P<0.05).4. The releationship between the expression of inflammatory cytokines and the airway and pulmonary vascular remodeling:There was positive correlation between the expressions of inflammatory cytokines (NF-κB,HMGB1) and airway remodeling index (MT%,MA%) and pulmonary vascular remodeling index (WT%,WA%) and inflammatory cytokines(IL-8,TNF-a,VEGF). There was positive correlation between the expressions of NF-κB and HMGB1. After using PDTC, the expression of inflammatory cytokines (NF-κB,HMGB1,IL-8, TNF-a,VEGF) were decreased and the airway remodeling and pulmonary vascular remodeling were improve partly.Conclusions:①With the development of the desease, the expression of TNF-α、 IL-8、VEGF in Serum and bronchoalveolar lavage fluid were gradually increased, there was positive correlation between the expressions of TNF-a, IL-8, VEGF and NF-κB,HMGB1. Prompted the inflammation of COPD is not only confined to the lungs, while there is a systemic inflammatory response. Systemic inflammatory response participated in all process of the development of COPD, Systemic inflammatory response expression advances gradually with the aggravation of COPD. It was possibly promotes the COPD course progress through the airway and pulmonary vascular remodeling.②NF-κB influence pulmonary vascular and airway reconstruction by regulating the expression of HMGB1, TNF-alpha, IL-8, VEGF. NF-κB plays an important role in COPD inflammation factor network.