Protective Effect And Mechanism Of Zhuyang Bufei Chubi Granule On Pulmonary Fibrosis Rats Based On Notch/Jagged Signaling Pathway

Objective: To observe the effects of the drug on pathological morphology,Notch receptor,ligand,Activase and related molecules in rats with pulmonary fibrosis.To study its effects on pulmonary inflammatory response,extracellular matrix metabolism,pulmonary microangiogenesis,immune response and oxidative stress and other pathophysiological processes.To further explore the pathogenesis of pulmonary fibrosis and provide new ideas for the treatment of pulmonary fibrosis.Methods: 60 male wstiar rats were randomly divided into blank group(n = 10),model group(n = 10),prednisone acetate group(n = 20)and Zhuyang Bufei Chubi Granule group(n = 20).From the 2nd day after the successful establishment of the model,the blank group and the model group were given orally with distilled water solution at a daily dose of 10 ml/kg per unit weight;the prednisone acetate group was given 1:1 volume of prednisone acetate distilled water solution at a daily dose of 405 ?g/kg;the Buyang Bufei Chubi Granule group was given Zhuyang Bufei Chubi Granule(adult dose of 45 g)on the 2nd day after the establishment of the model./ The equivalent dose for adults was 3.5g/kg.d and the concentration of the solution was 35%.Once a day,28 days of continuous treatment.On the 7th,14 th and 28 th day after administration,the animals died in batches and their lungs were taken out.The anesthetics used in the anesthesia of rats were chloral hydrate(10%)at a dose of 3.ml/kg.After taking blood samples from the carotid artery,the lungs were dissected and trachea and main bronchus were cut off.Pulmonary tissues were washed with ice-salt water for three times,then water was absorbed by filter paper,then fixed with formaldehyde solution(10%)to the left lung,and then paraffin-embedded sections were processed.The thickness of sections was about 4-5 um,then stained with HE and Masson.The remaining lung tissues were placed in EP tube(1.5 ml)and stored in ultra-low temperature refrigerator(-80 ?).Use.The ratio of target protein to ?-actin gray value was calculated by using the software ImageLab 4.0 in the step of Westenblot experiment.The ratio was used to express the expression level of target protein.All the data are represented in x±s form.Statistical processing is accomplished by software SPSS18.0,and multigroup comparisons are made by one-way ANOVA.Results: The expression levels of Notch1 and Jagged1 in blank group were lower than those in prednisone acetate group and Zhuyang Bufei Chubi Granule group(P < 0.05).Compared with model group,the expression levels of Notch1 and Jagged1 in prednisone acetate group and Zhuyang Bufei Chubi Granule group decreased significantly(P < 0.05).Compared with prednisone acetate group,the expression levels of Notch1 and Jagged1 in Zhuyang Bufei Chubi Granule group were higher(P < 0.05).Regulation of EMT in lungs showed that Zhuyang Bufei Chubi Granule had a protective effect on Bleomycin-induced pulmonary fibrosis in rats.Conclusion: In this experiment,the effects of Zhuyang Bufei Chubi Granule on the pathological characteristics,Activase and its related proteins,Notch receptor and ligand of pulmonary fibrosis rat model were observed: Zhuyang Bufei Chubi Granule could reduce the degree of pulmonary fibrosis,reduce the expression of Notch1,Jagged1,CD54,ADAM17,VCAM-1 protein in model group and increase the expression of peroxiredoxin3 protein in model group.Zhuyang Bufei Chubi Granule can inhibit Bleomycin-induced pulmonary fibrosis and prevent the progress of pulmonary fibrosis.