Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology, molecular docking, and in vitro experiments

To explore and validate the potential targets of Paeoniae Radix Alba (P. Radix, Bai Shao) in protecting against chemical liver injury through network pharmacology, molecular docking technology, and in vitro cell experiments. Network pharmacology was used to identify the common potential targets of P. Radix and chemical liver injury. Molecular docking was used to fit the components, which were subsequently verified in vitro. A cell model of hepatic fibrosis was established by activating hepatic stellate cell (HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1. The cells were treated with various concentrations of total glucosides of paeony (TGP), the active substance of P. Radix, and then their viability was assessed using the Cell Counting Kit-8 assay. The expression levels of heat shock protein (HSP)-90α, prostaglandin endoperoxide synthase 2 (PTGS2), interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor α (TNF-α) were measured using enzyme-linked immunosorbent assay, whereas the protein expression levels of collagen type I (COL-I), collagen type Ⅲ (COL-III), peroxisome proliferator-activated receptor (PPAR)-γ, and caspase 3 (CASP3) were detected using western blot. Analysis through network pharmacology revealed 13 key compounds of P. Radix, and the potential targets for preventing chemical liver injury were IL-6, AKT serine/threonine kinase 1, jun proto-oncogene, heat shock protein 90 alpha family class A member 1 (HSP90AA1), peroxisome proliferator activated receptor gamma (PPARG), PTGS2, and CASP3. Gene Ontology (GO) enrichment analysis indicated the involvement of response to drugs, membrane rafts, and peptide binding. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation. Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1, PTGS2, PPARG, and CASP3. Different concentrations of TGP can inhibit the expression of COL-Ⅰ, COL-Ⅲ, IL-6, TNF-α, IL-1β, HSP-90α, and PTGS2 while increasing the expression of PPAR-γ and CASP3 in activated HSC-LX2 cells. The protective effect of P. Radix against chemical liver injury involves multiple components, targets, and pathways. P. Radix primarily functions by regulating targets such as HSP90AA1, PTGS2, PPARG, CASP3, and others. TGP, the main active compound of P. Radix, protects against chemical liver injury by reducing the inflammatory response, activating apoptotic proteins, and promoting the apoptosis of activated HSCs. In the future, the mechanism of action of P. Radix against chemical liver injury can be studied from two perspectives: regulating inflammatory response and apoptosis.