An integrated network pharmacology and proteomics approach reveals the anti-fibrotic effect of Fushen Granule on peritoneal fibrosis

Fushen Granule (FSG), a Chinese medicine formular, was used clinically to improve the efficiency of dialysis in end-stage renal disease patients receiving peritoneal dialysis treatment. However, the mechanisms of its antifibrotic effect on peritoneal fibrosis (PF) have not yet been studied. In this study, we aimed to identify the potential mechanism of FSG in intervening in PF. Network pharmacology analysis and molecular docking were used to predict the related ingredients and potential targets of FSG in treating PF. TGF-β1 induced MeT5A cells were used for in vitro verification. Label-free proteomics analysis was performed to detect the differentially expressed proteins in TGF-β1 induced MeT5A cells upon treatment with FSG-containing serum. Western blot and immunofluorescence assays were used to evaluate the expression of proteins related to PF and PI3K/AKT signalling. PPI network analysis showed that TP53, EGFR, HSP90AA1, AKT1, CCND1, MYC, STAT3, SRC and ESR1 constitute the core network. KEGG analysis found that the PI3K/AKT signalling pathway was significantly enriched. Molecular docking analysis indicated that the main active ingredient from the component–target network can closely integrate with targets in the PI3K/AKT signalling pathway. Proteomics analysis on TGF-β1-treated MeT5A also indicated that the PI3K/AKT signalling pathway was closely correlated with the effect of FSG in intervening PF. Further in vitro experimental validation showed that FSG decreased the protein levels of FN, Col Ⅰ, α-SMA, ITGβ3, PDK1, p-mTOR and p-AKT and reversed the expression of E-cad in TGF-β1-treated MeT5A cells. FSG showed therapeutic effects against PF, mainly by inactivating PI3K/AKT signalling.