Abstract： Objective： To explore the main active components and potential targets and pathways of Jianpi Shugan Qinghua Prescription by network pharmacology and in vitro experiments，and to preliminarily explore the mechanism of Jianpi Shugan Qinghua Prescription in treating non-alcoholic fatty liver disease (NAFLD). Methods： The main components and targets in Chinese medicinals in Jianpi Shugan Qinghua Prescription were screened in TCMSP database，and NAFLD related targets were screened in GeneCards， OMIM and TTD databases. After the intersection of the above two，the potential targets of Jianpi Shugan Qinghua Prescription in treating NAFLD were obtained，and the network diagram was drawn by Cytoscape. Protein interaction nework diogram was obtained by String database， and core genes were searched by Cytoscape. GO and KEGG enrichment analysis was carried out via the clusterProfiler package of R language. AutoDock Tools was used to simulate the interaction between target proteins and active components. The AML12 steatosis cell model was constructed with palmitic acid and oleic acid. The therapeutic effect and mechanism of Jianpi Shugan Qinghua Prescription in treating NAFLD were verified by cell oil red O staining， intracellular glycerin trilaurate (TG) determination and mRNA relative levels of core targets. Results：1) After oral bioavailability (OB) and drug-likeness (DL) screening， 210 kinds of active components were found in 11 kinds of Chinese medicinals，among which quercetin，kaempferol，luteolin，isorhamnotin，β- sitosterol and naringin were the main active components. 2) There were 119 potential targets in Jianpi Shugan Qinghua Prescription in treating NAFLD，among which TNF，AKT1，IL-6，IL-1β，PPARG and TP53 were the core targets. 3) GO analysis showed that the regulation of reactive oxygen species metabolism (GO： 2000377)， response to lipopolysaccharide (GO： 0032496)， reactive oxygen species metabolism (GO： 0072593)， response to bacteriological molecules (GO： 0002237) and response to drugs (GO： 0042493) played a key role. KEGG enrichment analysis showed that tumor necrosis factor signaling pathway，NAFLD， toll-like receptor pathway， insulin resistance pathway and other pathways might play a key role in treatment. 4) Molecular docking showed that quercetin and IL-1β were well bound. 5) In vitro cell experiments showed that Jianpi Shugan Qinghua Prescription could reduce the area of oil red staining (lipid stain) and intracellular TG levels in liver cells， and RT-qPCR showed that Jianpi Shugan Qinghua Prescription could regulate the expression levels of core targets. Conclusion： Based on network pharmacology and in vitro experiments， the mechanism of Jianpi Shugan Qinghua Prescription in treating NAFLD may be due to the effect of quercetin， kaempferol， luteolin on TNF， AKT1， IL-6， IL-1β， PPARG， TP53 and other targets through TNF signaling pathway， toll-like receptor pathway， and insulin resistance pathway. It is the result of multi-component，multi-target and multi-path interaction.