Integration of Network Pharmacology Analysis Investigating the Functional Role of the p53 Signaling Pathway in the Improvement of Pulmonary Fibrosis in A Mouse Model by Maimendong Decoction

LI Jie; KONG Xinyi; ZOU Jun; LI Jinghong

Abstract：Objective To investigate the interventional effects of Maimendong decoction on a mouse model of pulmonary fibrosis and the underlying pharmacological mechanisms. Methods C57BL/6 mice were randomly allocated into three groups:control group,model group,and treatment group,with 6 mice in each group. A pulmonary fibrosis mouse model was established by tracheal instillation of bleomycin in the model group,and 24 hours after the model was established,the intervention group was given Maimendong decoction by gavage for 20 days. The pulmonary fibrosis mouse model was established by tracheal instillation of bleomycin in the model group. Twenty-four hours after the model was established,mice in the treatment group were given Maimendong decoction by gavage for 20 days. The general condition of the mice was observed,and the pathological alterations in lung tissue were examined by HE staining. The expression of alpha smooth muscle actin (α-SMA) and epithelial cadherin (E-cadherin) was analyzed by immunohistochemistry staining and Immunoblotting. Network pharmacology analysis was performed to explore the potential mechanism of Maimendong decoction in improving pulmonary fibrosis. Furthermore,a pulmonary fibroblast model was established by inducing mice embryonic fibroblast NIH3T3 with transforming growth factor-β1 (TGF-β1). After treatment with Maimendong Decoction-containing serum and/or p53 inhibitor (Pifithrin-α,PFT-α),the expression of p53,α-SMA,E-cadherin,and collagen type I (Collagen I) was detected by real-time PCR and Immunoblotting. Results Compared with the control group,the lung tissues of mice in the pulmonary fibrosis model group showed extensive consolidation areas,abundant infiltration of inflammatory cells,increased expression of α-SMA,and decreased expression of E-cadherin (P<0. 01). These features were significantly improved after Maimendong decoction treatment. Network pharmacological analysis revealed that the differentially targeted genes of Maimendong decoction were mainly enriched in the p53 signaling pathway. Cell experiments showed that the treatment with Maimendong decoction-containing serum led to the downregulation of α-SMA and upregulation of E-cadherin and p53 in the pulmonary fibroblast model (P<0.01). In the PFT-α treatment experiment,the expression of p53 and E-cadherin was reduced in both the control and Maimendong decoction groups,while the expression of α-SMA and Collagen I was increased (P<0. 01). Conclusion Maimendong decoction can improve the progression of pulmonary fibrosis by regulating the p53 signaling pathway.