| Objective: This topic is based on warming the lungs,invigorating qi and dredging collaterals,and taking the Nrf2/HO-1 signal pathway as the entry point,and the effect of Congfei Buqi Tongluo Recipe on the pathological changes in serum or lung tissue of mice with pulmonary fibrosis,and the influence of inflammatory factors TGF-β1 and IL-1β,Antioxidant-related factors ROS,antioxidant enzymes SOD,GSH-px,lipid peroxi-dation product MDA,and Nrf2,HO-1,Keap1 protein expression changes in four aspects to conduct experimental research.To confirm the mechanism of Wenfei Buqi Tongluo Decoc-tion on bleomycin-induced pulmonary fibrosis in mice,provide experimental ideas for the clinical application of Wenfei Buqi Tongluo Decoction in the treatment of interstitial lung diseases,and lay a theory for clinical research basis.Methods:1.Effect of Wenfei Buqi Tongluo Recipe on the pathological morphology of lung tissue in mice with pulmonary fibrosis induced by bleomycin.The pulmonary fibrosis model of mice was established by intratracheal instillation of bleomycin.The general condition,overall morphology,he staining,Masson method and sazpiel score of mice were observed dynamically,and the effect of Wenfei Buqi Tongluo Recipe on lung histopathology of mice with pulmonary fibrosis was observed.2.Effect of Wenfei Buqi Tongluo Recipe on IL-1 β and TGF-β 1 in serum of mice with pulmonary fibrosis.Pulmonary fibrosis mice were collected on the 7th,14 th and 28 th day after drug intervention.The serum levels of IL-1 β and TGF-β 1 were detected by ELISA.3.Effect of Wenfei Buqi Tongluo Recipe on antioxidant factors in lung tissue of mice with pulmonary fibrosis.The contents of ROS,SOD,GSH-px and MDA in lung tissue of mice with pulmonary fibrosis were detected by ELISA.4.Effect of Wenfei Buqi Tongluo Recipe on the expression of Nrf2,HO-1 and Keap1 in lung tissue of mice with pulmonary fibrosis.Western-blot method was used to detect the expression of Nrf2,HO-1,Keap1 protein in the lung tissues of each group of mice after drug intervention.Results:1.Effect of Wenfei Buqi Tongluo Recipe on Pathomorphology of lung tissue in mice with bleomycin induced pulmonary fibrosis(1)He staining: Compared with the model group,alveolitis degree of lung tissue in the posit-ive drug group and the Chinese medicine group were improved in different degrees compared with the model group(P<0.001).(2)Masson staining: The results were most obvious on the 14 th day.In the model group,alveolar septum widened,interstitial fibroblasts increased,collagen fibers increased and diffusely distributed,and serious collagen deposition appeared.Compared with the model group,the fibroblasts and collagen deposition in the lung interstitium of the positive drug group and the high and medium dose Wenfei Buqi Tongluo Recipe groups were significantly improved(P<0.001).2.Effect of Wenfei Buqi Tongluo Recipe on IL-1β and TGF-β 1in serum of mice with pulmonary fibrosis(1)IL-1β: Compared with the blank group,the levels of IL-1β in the serum of mice in each group increased to varying degrees(P<0.01),especially in the model control group(P<0.001).Compared with the model group,the serum IL-1β levels of mice in each administration group decreased to different degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo Recipe.The high-dose group decreased significantly(P<0.001),especially on the 14 th day(P<0.001).(2)TGF-β1: Compared with the blank group,the levels of TGF-β1 in the serum of mice in each group increased to varying degrees(P<0.01),especially in the model control group(P<0.001).Compared with the model group,the serum TGF-β1 content of mice in each administration group decreased to different degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo prescrip-tion.The high-dose group decreased significantly(P<0.001),especially on the 14 th day(P<0.001).3.Effect of Wenfei Buqi Tongluo Recipe on antioxidant factors in lung tissue of mice with pulmonary fibrosis.(1)ROS: Compared with the blank group,the content of ROS in the serum of mice in each group increased to varying degrees(P<0.01),especially in the model control group(P<0.001).Compared with the model control group,the content of ROS in the lung tissue of mice in each treatment group at different time periods was up-regulated in varying degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo Recipe middle dose group and high dose group(P<0.001).(2)SOD: Compared with the blank group,the SOD content in the serum of mice in each group decreased to varying degrees(P<0.01),especially in the model control group(P<0.001).Compared with the model control group,the content of SOD in the lung tissue of mice in each treatment group at different time periods was up-regulated in varying degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo Recipe middle dose group and high dose group(P<0.001).(3)GSH-px: Compared with the blank group,the content of GSH-px in the serum of mice in each group decreased to varying degrees(P<0.01),especially in the model control group(P<0.001).Compared with the model control group,the content of GSH-px in the lung tissue of mice in each treatment group at different time periods was up-regulated in varying degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo Recipe middle dose group and high dose group(P<0.001).(4)MDA:Compared with the blank group,the levels of MDA in the serum of mice in each group increased to varying degrees(P<0.01),especially in the model control group(P<0.001).Compared with the model control group,the content of MDA in the lung tissue of mice with in each administration group was down-regulated to varying degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo Recipe The dose group and high dose group were down-regulated significantly(P<0.001).4.Effect of Wenfei Buqi Tongluo Recipe on the expression of Nrf2,HO-1 and Keap1 in lung tissue of mice with pulmonary fibrosis.(1)Nrf2: Compared with the model control group,the protein expression levels of mice in the positive drug group and Wenfei Buqi Tongluo Decoction groups were increased in varying degrees(P<0.01),especially in the positive drug group and Wenfei Buqi Tongluo Decoction high-dose group(P<0.001);compared with the posit-ive drug group,the Nrf2 protein level of Wenfei Buqi Tongluo Decoction high-dose group was not increased The difference was significant(P>0.05).(2)HO-1: Compared with the model control group,the protein expression levels in the lung tissues of mice in the positive drug group and the various dose groups of Wenfei Buqi Tongluo Recipe were up-regulated to varying degrees(P<0.01),especially in the positive drug group and The high-dose Wenfei Buqi Tongluo Recipe group was significantly up-regulated(P<0.001);compared with the positive drug group,the HO-1 protein level of the high-dose Wenfei Buqi Tongluo Recipe group had no significant difference(P>0.05).(3)Keap1:Compared with the model control group,the protein expression levels of the lung tissues of mice in the positive drug group and the various dose groups of Wenfei Buqi Tongluo Recipe were down-regulated to varying degrees(P<0.01),especially the positive drug group and The high-dose Wenfei Buqi Tongluo Recipe group was significantly down-regulated(P<0.001);compared with the positive drug group,there was no significant difference in the degree of Keap1 protein level down-regul-ation in the high-dose Wenfei Buqi Tongluo Recipe group(P>0.05).Conclusion:Based on the above experimental results,it is concluded that the effects of Wenfei Buqi Tongluo Recipe on pulmonary fibrosis have the following 4 conclusions:1.It can significantly reduce the formation of alveolitis and pulmonary fibrosis in lung tissues of mice with pulmonary fibrosis,and its effect is similar to that of pirfenidone.2.By regulating the oxidative stress response,down-regulate the content of inflammatory factors IL-1β and TGF-β1,improve alveolitis,and inhibit the formation of pulmonary fibrosis.3.By reducing the content of ROS and MDA and up-regulating the content of SOD and GSH-px,especially in the high-dose group,it can regulate the balance of antioxidant system and inhibit the formation of pulmonary fibrosis.4.By regulating the antioxidant Nrf2/HO-1 signal pathway,down-regulating the protein expression of Nrf2 and HO-1,thereby delaying the progression of pulmonary fibrosis. |
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