To Explore The Inflammatory Mechanism Of Qingfehuatan Decoction In Chronic Obstructive Pulmonary Disease From P38MAPK Signaling Pathway

Objective:To establish a rat model of chronic obstructive pulmonary disease(COPD),and to explore the inflammatory mechanism of Qingfehuatan Decoction on COPD from the p38 MAPK pathway,and to further explore the therapeutic mechanism of Qingfehuatan Decoction on COPD.Methods:Sixty healthy SPF Wistar rats,male,with the body weight of(200±20)g,were randomly divided into 6 groups according to the random number table method: normal group(group A,n=10),model group(group B,n=10),Qingfei Huatan Decoction low-dose group(group C,n=10),Qingfei Huatan Decoction medium-dose group(group D,n=10),qingfei Huatan decoction low-dose group(group C,n=10),And Qingfei Huatan Decoction medium-dose group(group D,n=10).Qingfehuatan Decoction high-dose group(group E,n=10),roxithromycin group(group F,n=10).The COPD model rats were established by smoking combined with trachea injection of lipopolysaccharide(LPS)and treated with Qingfehuatan Decoction.The effect of Qingfehuatan Decoction on COPD was observed by HE staining.The contents of IL-6,TNF-α and CCL5 in serum and alveolar lavage fluid(BALF)were determined by enzymic linked immunosorbent assay(ELISA).The expression level of p38 MAPK m RNA in lung tissues was detected by immunofluorescence PCR.Western Blot(WB)was used to detect the relative expression of p38 MAPK protein.Results:(1)Pathological changes of lung tissue: Model group is a large amount of inflammatory cells infiltration in the lung tissue,give priority to with lymphocytes and macrophages,bronchial wall thickening of the intima,luminal stenosis,mucosal epithelial cell necrosis,fall off,the cilia arranged disorder,airway submucosal glands hyperplasia,goblet cells increased,leading to airway mucus secretion increased,the visible part of the tube cavity with sputum bolt formation,alveolar wall thinning,alveolar space greatens,Partial fusion to form pulmonary bullae suggests successful modeling.The damage degree of lung tissue in qingfehuatan decoction low-dose,medium-dose and high-dose groups and roxithromycin group was reduced.(2)Changes of IL-6,TNF-α and CCL5 contents in serum and BALF of rats in each group: Compared with blank group,the contents of IL-6,TNF-α and CCL5 in serum and BALF of rats in model group were significantly increased(P<0.05);Compared with model group,the contents of IL-6,TNF-α and CCL5 in serum and BALF of Qingfehuatan Decoction low-dose,medium-dose and high-dose groups and roxithromycin group were decreased,with statistical significance(P<0.05).The contents of IL-6,TNF-α and CCL5 in serum and BALF of qingfehuatan Decoction low-dose,medium-dose and high-dose groups decreased more obviously with the increase of drug dose,and the difference was statistically significant(P<0.05).Both qingfehuatan decoction high-dose group and roxithromycin group could significantly reduce the level of inflammatory factors in rats,and there was no statistical significance between them(P>0.05).(3)Changes of p38 MAPK m RNA and p38 MAPK protein expression in lung tissues of rats in each group: compared with normal group,p38 MAPK m RNA and p38 MAPK protein expression in lung tissues of model group were significantly increased,and the difference was statistically significant(P<0.05);Compared with model group,the expression of p38 MAPK m RNA and p38 MAPK in lung tissues of rats in qingfei Huatan Decoction low-dose,medium-dose and high-dose groups and roxithromycin group were significantly decreased(P<0.05),and the decrease was more obvious in Qingfei Huatan Decoction high-dose group and roxithromycin group,and there was no statistical significance between them(P>0.05).Conclusion:(1)The COPD rat model can be successfully replicated by smoking combined with tracheal injection of lipopolysaccharide.(2)Qingfei Huatan Decoction can significantly improve the symptoms of COPD model rats and reduce pathological damage.(3)Qingfei Huatan Decoction can reduce airway inflammation in COPD rats and play a protective role in lung tissue.The mechanism may be to reduce the release of inflammatory factors such as IL-6,TNF-α and CCL5 by down-regulating the over-expression and continuous activation of p38 MAPK signaling pathway,so as to reduce the inflammatory response and achieve the effect of treating COPD.