Objective. The mechanism of action of asiatic acid (AA) on alcoholic steatohepatitis (ASH) was investigated using network pharmacology and experiments. Methods. Through data retrieval, network construction, and enrichment analysis, the potential mechanism of AA in the treatment of alcoholic steatohepatitis was explored. Animal and cell models were established in this study. Animal Model. The mouse model was divided into six groups: normal group; model group; low, medium, and high AA group; and silibinin-positive group. Cell Model. An in vitro inflammatory model of RAW264.7 cells was established by alcohol stimulation. Results. Compared with the model group, the low, medium, and high AA group showed decreased serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and total cholesterol (T-CHO). The inflammatory factor tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6) in a dose-dependent manner were decreased. In addition, hematoxylin-eosin staining showed that liver tissue damage and inflammatory cell infiltration in mice were significantly reduced with increasing doses. Further, oil red staining showed that lipid accumulation in hepatocytes in the low, medium, and high AA group was significantly reduced, with increasing dose. In addition, in the cellular model, real-time reverse transcriptase-polymerase chain reaction (Real-Time RT-PCR) and enzyme-linked immunosorbent assay (ELISA) results showed that AA could alleviate alcohol-induced cellular inflammation, while western blot and immunofluorescence results showed that AA might alleviate alcohol-induced cellular inflammation by inhibiting the nuclear factor-κB (NF-κB) pathway. Conclusion. This study provides multiple lines of evidence that asiatic acid may alleviate alcoholic hepatitis in mice by modulating the NF-κB pathway.
Purpose: This study utilized network pharmacology, molecular docking, molecular dynamics simulation, and in vitro experimental verification to explore the mechanisms of action by which asiatic acid (AA) inhibits acetaldehyde-induced activation of hepatic stellate cells (HSCs). Methods: Databases were screened to identify intersections between AA and alcoholic liver fibrosis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment analysis were performed, followed by molecular docking and molecular dynamics simulation to predict further the interactions between AA and cross-targets. Mechanisms of action of AA were assessed experimentally in HSC-T6 rat HSCs. Results: Screening of the relevant databases using web pharmacology identified several genes, including those encoding signal transducer and activator of transcription 3, nuclear factor kappa-B (NF-κB)-P65 (RELA), tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 1 beta (IL-1β), that were closely associated with alcoholic liver fibrosis. GO and KEGG enrichment analysis showed that the alcoholic liver disease pathway was the most relevant one, and molecular docking showed critical binding activity of RELA, TNF-α, IL-6, and IL-1β with AA. Molecular dynamics simulation showed that the binding between RELA and AA was stable. AA at concentrations of 0 to 30 μM were determined to be nontoxic to HSC-T6 cells. Real-time quantitative polymerase chain reaction and Western blotting showed that the levels of expression of α-SMA and type 1 collagen were higher in acetaldehyde-treated than untreated HSC-T6 cells, with AA reducing their expression dose-dependently in acetaldehyde-treated cells. Western blotting also showed that AA could upregulate the expression of the apoptotic proteins BCL2 associated X and cleaved caspase 3 and could inhibit the phosphorylation of RELA and IKB-α. Conclusion: AA inhibited acetaldehyde-induced HSC-T6 cell proliferation and suppressed the secretion of fibrillar factors by inhibiting RELA phosphorylation.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.