Elucidating the Key Bioactive Constituents and Preventive Mechanism of Sea Buckthorn Sterols against Non-Alcoholic Fatty Liver Disease

This study employed an integrated approach combining network pharmacology, molecular docking, and experimental validation to systematically explore the therapeutic basis and underlying mechanisms of sea buckthorn sterols in the treatment of non-alcoholic fatty liver disease (NAFLD). Potential targets of sea buckthorn sterols were predicted using the Swiss Target Prediction, TargetNet, and SEA databases, while NAFLD-related targets were retrieved from DisGeNET, GeneCards, and OMIM databases, enabling the construction of a compound-target-disease network. Functional enrichment analysis revealed key biological processes and signaling pathways implicated in the intervention. Key bioactive components were subsequently subjected to molecular docking studies using AutoDock to evaluate their binding potential with key targets. In vitro experiments were conducted using free fatty acid-induced HepG2 cells to assess the effects of sea buckthorn sterols on lipid metabolism by quantifying total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) levels, with intracellular lipid accumulation visualized via Oil Red O staining. The results identified 86 overlapping targets between sea buckthorn sterols and NAFLD, including 10 key targets such as PPARG, AKT1, and ESR1, along with 17 key sterol components, including cerevisterol, lanosterol, and delta7-stigmasterol. KEGG pathway analysis indicated involvement in 130 signaling pathways, with prominent roles in insulin resistance, HIF-1 signaling pathway and C-type lectin receptor signaling pathway. Molecular docking confirmed stable binding interactions between the key sterol components and their respective targets. Cellular assays demonstrated that sea buckthorn sterols significantly reduced TC, TG, and LDL levels, increased HDL level, and markedly attenuated intracellular lipid droplet accumulation. In conclusion, cerevisterol and delta7-stigmasterol, as principal active constituents of sea buckthorn sterols, likely ameliorate NAFLD by modulating lipid metabolism, insulin resistance, and glucose homeostasis through multi-target interactions involving PPARG, AKT1 and ESR1.