Investigation Of Tcm Syndromes Of AE-IPF And The Effect Of Bushen Huoxue Method On TGF-?1/Smads Signal Pathway In Rats With Pulmonary Fibrosis

Objective1Clinical Research: To explore the distribution of TCM syndromes in acute exacerbation of idiopathic pulmonary fibrosis(AE-IPF).2 Animal experiment:To explore the effect of Bushen Huoxue Method on TGF-?1/Smads signal pathway in rats with pulmonary fibrosis.Methods1 Clinical Research:Collect relevant data and develop AE-IPF TCM syndrome questionnaire.In the form of questionnaire,the relevant data of patients and the four diagnostic information of traditional Chinese medicine were collected,collated and analyzed to explore the distribution characteristics of TCM syndrome types of patients with AE-IPF.2 Animal experiment: SD rats were randomly divided into blank group,model group,prednisone group,Bushen Huoxue capsule low,middle and high dose groups.The model of pulmonary fibrosis was induced by intratracheal injection of bleomycin,and the samples were taken on the 28 th day after administration.After the rats were killed,the pathology of lung tissue was observed by hematoxylin-eosin staining(HE)and Masson(Masson)trichromatic staining,the expression of HYP and KL-6 in lung tissue was determined by immunohistochemical technique,and the expression of TGF-?1,TNF-? and LTB4,in serum was detected by ELISA method.The expression of Smad2/3,Smad7 and p-Smad2/3 protein in lung tissue was detected by Western Blot.Results1 Clinical studies:The common TCM syndromes of 60 AE-IPF patients were phlegm-dampness stasis lung syndrome and lung-kidney qi-yang deficiency syndrome(18.33%),phlegm-heat stasis lung syndrome and lung-kidney qi-yang deficiency syndrome(13.33%),phlegm-heat stasis lung syndrome and lung-qi-yin deficiency syndrome(10%),phlegm-dampness blocking lung syndrome and lung-kidney qi-yang deficiency syndrome(8.33%),phlegm-heat stasis lung syndrome and lung-kidney yin deficiency syndrome(8.33%).Among all the syndrome types,blood stasis syndrome accounted for 63.33%,and those without blood stasis syndrome accounted for 36.67%.2 Animal experiment2.1 HE staining: The outline of lung tissue structure of rats in the blank group was basically complete and clear.In the model group,alveolar structure destruction,local inflammatory cell infiltration,alveolar septum thickening,a large number of fibroblasts proliferation and fibrous tissue could be seen.The pathological changes of lung tissue of rats in each treatment group were alleviated in varying degrees,among which the high dose Bushen Huoxue capsule group and prednisone group showed the most obvious reduction,followed by low and middle doses.2.2 Masson staining:A small amount of blue-stained collagen fibers were distributed in the alveolar septum of rats in the blank group.Dark blue collagen fibers were observed in the model group,mainly in the terminal bronchial wall,vascular wall and alveolar septum.Compared with the model group,the lung tissue of each treatment group became lighter,the thickness of alveolar septum was narrower,and the degree of fibrosis was alleviated.The improvement of high dose Bushen Huoxue capsule group and prednisone group was the most obvious,and the collagen deposition was the least.2.3 The expression of HYP and KL-6:Compared with the blank group,the expression of HYP and KL-6 in the lung tissue of the other groups was significantly higher,and the expression of HYP and KL-6 in the model group was the highest,while the expression of HYP and KL-6 in each treatment group was lower than that in the model group,and the difference was statistically significant between the high dose Bushen Huoxue capsule group and the prednisone group(P<0.05).2.4 The contents of TGF-?1?TNF-??LTB4:Compared with the blank group,the contents of TGF-?1,TNF-? and LTB4 in the serum of the other groups were significantly higher,and the contents of TGF-?1,TNF-? and LTB4 in the model group were the highest,while the contents of TGF-?1,TNF-? and LTB4 in each treatment group were lower than those in the model group,and the difference was statistically significant(P<0.01).Compared with each treatment group,the contents of TGF-?1,TNF-? and LTB4 in serum in high dose Bushen Huoxue capsule group and prednisone group decreased most significantly,and the difference was statistically significant(P<0.01).2.5 The expression of Smad2/3,p-Smad2/3 and Smad7 protein:Compared with the blank group,the expression of Smad2/3 and p-Smad2/3 protein in lung tissue of rats in other groups increased significantly,while the expression of Smad7 protein decreased significantly,and the difference was statistically significant(P<0.05).Compared with the model group,the expression of Smad2/3 and p-Smad2/3 protein in lung tissue of rats in each treatment group decreased significantly,while Smad7 increased significantly,and the difference was statistically significant(P<0.05),among which the decrease of Smad2/3 and p-Smad2/3 and the increase of Smad7 in high dose Bushen Huoxue capsule group and prednisone group were the most significant(P<0.05).Conclusion1 The acute exacerbation of idiopathic pulmonary fibrosis is characterized by mixed syndrome of deficiency and excess.2 Bushen Huoxue capsule can inhibit collagen deposition,reduce the proliferation of type II alveolar epithelial cells,reduce pulmonary inflammation,reduce cytotoxicity and regulate the balance of cytokines by reducing the contents of TNF-? and LTB4,so as to reduce the injury of lung tissue induced by bleomycin in rats.3 Bushen Huoxue capsule can reduce the serum content of TGF-?1,inhibit the expression of Smad2/3 and p-Smad2/3 protein in lung tissue,and promote the expression of Smad7 protein.It is speculated that Bushen Huoxue capsule can inhibit pulmonary fibrosis by regulating TGF-?1/Smads signal pathway.