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：A total of 16 C57BL/6J mice aged 6-weeks old were randomly allocated into control group and NASH group（control n=8；NASH n=8）. Control mice were fed with a standard chow diet and water as control group，while the other mice were fed high fat diet accompanied by ad libitum consumption of water with 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 HE 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. Results：①The levels of alanine aminotransferase（ALT） and aspartate aminotransferase（AST） in NASH mice were significantly higher than that in control group（P < 0.05）. ②Compared 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. Conversely，total ceramide and total lactosylceramide（LacCer） were remarkably elevated in NASH group than control group（P < 0.05）. Conclusion：The synthetic pathway from LacCer to GM3 is dampened in NASH model mice，which are established by high-fat and high-fructose diet.