The potential mechanisms of Saikosaponin A for Non-Alcoholic Fatty Liver Disease: An integration of network pharmacology, molecular docking, molecular dynamics simulationand experimental validation

Non-alcoholic fatty liver disease (NAFLD) is a leading cause of liver disorders globally, which is affecting the health of over one-third of the global population. Our study aimed to investigate potential mechanisms and drug targets of Saikosaponin A (SSA) in treating NAFLD.We explored the effects of SSA on NAFLD through a combination of network pharmacology, molecular docking, molecular dynamics simulations, and in vitro experiments using oleic acid (OA) and palmitic acid (PA)-treated mouse primary hepatocytes. Network pharmacological analysis showed 25 intersecting genes were obtained by venny analysis. GO and KEGG analysis showed 386 biological process, 13 cellular components, 24 molecular function and 90 signaling pathways were involved. The results showed that SSA may improve the lipid metabolism in NAFLD, and apoptosis may the most significant mechanism as P53 was the key gene among hub genes, which are involved in P53/BCL2 pathway.In addition, oxidative stress may another important mechanism which further validation is required. Importantly, molecular docking and molecular dynamics simulation showed that P53 and Nrf2 could interacts stably with SSA, and WB, IF and qPCR validates the convincible change of P53/BCL2/CASP3 and Nrf2/HO-1 pathway.The findings suggest that SSA can significantly improve apoptosis and oxidative stress in NAFLD, in which, the underlying mechanism are associated with the P53/BCL2 and Nrf2/HO-1 pathway.