Hepatic gene expression in CDAHFD-induced non-alcoholic steatohepatitis model mouse

A choline-deficient amino acid defined high-fat diet (CDAHFD)-induced NASH model is a well-established and reproducible experimental animal model. However, the cause of hepatic fibrogenesis on this model remains unclear. This study aimed to clarify the phenotypic changes and causality between metabolic changes with hepatic fibrogenesis in the CDAHFD-fed mice. To characterize the phenotypic changes in the CDAHFD-fed mouse liver, we conducted transcriptomic analyses (RNA-seq). C57BL/6J male mice were given a choline-deficient L-amino acid defined (0.1% methionine) 60% kcal high-fat diet (CDAHFD; A06071302, Research Diets) for 0, 1, 4, 8, and 12 weeks. The age- and sex-matched acclimatized mice fed a standard chow diet (CRF-1; Oriental Yeast, Japan) were assigned to the control group (0 weeks). Total RNA was extracted from frozen liver samples using the RNeasy Mini kit (Qiagen). After RNA quantification and quality control, libraries were constructed using the TruSeq Stranded mRNA LT Sample Prep kit (Illumina). The NovaSeq 6000 (Illumina) sequence paired-end read length was set to 2 × 100 bp; raw data were processed and checked using the Galaxy system. The filtered data were mapped onto the Gencode reference genome (GRCm38.p6) and annotated using Gencode (M24).