Objective To investigate the expression levels of hepatic immune-related genes in C57BL/6J mice treated by high-fat diet. Methods Male C57BL/6J mice were treated by high-fat diet for 16 and 38 weeks, and the control group were given normal diet. The body mass and blood glucose levels were measured, and the liver slices were checked by HE staining. Hepatic RNA was extracted and sequenced by a Beijing Genomics Institute (BGI) BGISEQ-500 Sequencing Platform. Original sequence data were aligned and annotated referring to UCSC GRCm38/mm10 mouse genome. Volcano plots were used to profile the mRNA expression data matrix and find out the most significantly regulated hepatic immune-related genes. Real-time PCR was performed to validate the changes of selected genes. Results High-fat diet significantly increased mouse body mass and blood glucose levels, and HE staining showed the accumulation of lipid in the livers of mice treated by high-fat diet. Volcano plots of transcriptome data showed that, high-fat diet significantly changed the expression levels of 157 out of 795 hepatic immune-related genes at week 16. At week 38, 235 genes were significantly changed, and most genes showed an up-regulation trend. Among those significantly changed genes, six genes were selected to validate the data with a real-time PCR technology. At week 16, the gene expression levels of CD36 and PR domain containing 1 (PRDM1) significantly increased, and CD276 significantly decreased. At week 38, the gene expression levels of chemokine CXC motif ligand 1 (CXCL1), CD40 and formyl peptode receptor 1 (FPR1) were significantly raised. The real-time PCR data were generally in line with the trend of transcriptomic data. Conclusion Long-term treatment with high-fat diet significantly changes the gene expression levels of mouse hepatic immune-related genes.