Objective: This study aims to investigate the changes of glycosphingolipid, especially the ganglioside GM3, in the liver of NASH model mice induced by high-fat and high-sugar diet . Methods: 6-weeks old C57BL/6J mice were randomly devided into control group and NASH group (control n=8; NASH n=8). Mice fed a standard chow diet yinormal water as control group, while the other mice fed high fat diet accompanied by ad libitum consumption of water with a high fructose and glucose content (23.1 g/L d-fructose +18.9 g/L d-glucose) to establish NASH models as model group. Serum levels of ALT and AST were measured to evaluate the pathological changes of liver tissues. Liver histology was assessed using hematoxylin and eosin (H&E) stains in paraffin-embedded sections using standard commercially used methods. Liver lipidomics analysis was perfonned in an electrospray ionization mode using Exion UPLC-QTRAP 6500 PLUS (Sciex) LC/MS. The student’s t test and Mann-Whitney U test were used for statistical analysis. Result: 1)The levels of ALT and AST in NASH mice were significantly higher than that in control group(P<0.05). 2)Compared with the control group, total GM3 and subtype species, including GM3 d18:1/20:0 （P=0.005）、GM3 d18:1/22:0（P=0.004）、GM3 d18:1/24:0（P=0.004）and GM3 d18:1/26:0（P=0.042）were significantly lower in the model group, which were dignosed as NASH group based on the NAFLD activity score（NAS）. Conversely, total ceramide and total laccer were remarkably elevated in NASH group(P<0.05). Conclusion: The synthetic pathway from LacCer to GM3 was blocked in NASH model mice, which were established by high-fat and high-fructose diet，resuting total GM3，part of Long chain fatty acid-GM3 and Very long chain fatty acid-GM3 were significantly reduced in the liver of NASH model mice.