This study aims to investigate the mechanism of Kudingcha against non-alcoholic fatty liver disease (NAFLD).Network pharmacology was used to predict the active ingredients and potential targets of Kudingcha in the treatment of NAFLD.The protein-protein interaction (PPI) network of Kudingcha in the treatment of NAFLD was constructed.Furthermore,the DAVID database was conducted to explore GO and KEGG enrichment analysis.Importantly,experimental tests were performed to determine the effect of core ingredients of Kudingcha on palmitic acid (PA) -induced cell viability,lipid deposition,apoptosis level and the expression of core targets in HepG2 cells.Network pharmacological analyses yielded nine active components of Kudingcha such as quercetin (QE) and kaempferol (KMP),as well as 101 potential targets for the treatment of NAFLD.Meanwhile,a total of eight core targets were screened from PPI network,such as tumor protein p53 (TP53) and V-Rel avian reticuloendotheliosis viral oncogene homolog A (RELA).GO enrichment analysis showed that the targets mainly affected the process of response to xenobiotic stimulus,nutrient levels and oxidative stress.On the other hand,KEGG pathway analysis obtained the signaling pathways including lipid and atherosclerosis,as well as apoptosis pathways.In vitro studies indicated that QE and KMP incubation could significantly improve PA-induced cell viability suppression in HepG2 and AML-12 cells.Besides,QE and KMP might also obviously reduce PA-induced lipid deposition and the levels of total cholesterol,triglyceride as well as apoptosis in HepG2 cells.In addition,QE and KMP could also inhibit the expression of TP53 and RELA.In conclusion,the therapeutic effect of Kudingcha on NAFLD may be connected to the regulation of apoptosis pathway via TP53 and RELA.