The objective of this study was to determine the effect of varying undigested NDF at 240 h of in vitro fermentation (uNDF₂₄₀) and physically effective NDF (peNDF) content of fresh cow rations on hepatic metabolism and gene expression. Multiparous Holstein cows (n = 56) were fed a common prepartum ration beginning 28 d before expected parturition and assigned randomly at calving to one of 2 postpartum diets differing in content of uNDF₂₄₀ and peNDF. High fiber (HF; 35.3% NDF, 12.2% uNDF₂₄₀, 23.2% peNDF; n = 27) and low fiber (LF; 32.8% NDF, 9.5% uNDF₂₄₀, 21.6% peNDF; n = 29) treatment diets were formulated for equivalent metabolizable protein (110 g/kg DM) and starch (24.8% DM), with higher fiber levels achieved through the addition of straw. At 29 DIM, cows fed HF were switched to the LF diet and all cows were fed the LF diet through 42 DIM. Liver biopsies were obtained from a subset of 40 cows on d 7 ± 1.1 (mean ± SD) and 14 ± 1.0 postpartum, liver tissue was immediately frozen in liquid nitrogen. Liver samples were analyzed for glycogen and triglyceride content, and analyzed via RT-qPCR to evaluate mRNA expression of key metabolic enzymes. Data were analyzed by repeated measures ANOVA with random effect of cow within treatment and fixed effects of treatment, time, parity group, and all interactions. Cows fed the HF diet tended (P < 0.15) to have higher liver triglycerides at d 14 and lower overall liver glycogen than cows fed LF. Cows fed the LF diet tended (P < 0.15) to have lower gene expression of pyruvate carboxylase at d 14 and lower overall expression of carnitine palmitoyltransferase 1A compared with cows fed HF. Expression of 3-hydroxy-3-methylglutaryl-coA synthase 2 tended to differ (P = 0.12) by treatment and parity group such that 2nd lactation cows fed LF had lower expression than 3rd and greater lactation cows fed the LF diet, while cows fed HF had no difference due to parity. Cows fed the HF diet had altered liver metabolism in both glucose and fatty acid metabolic pathways, likely due to intake restrictions beginning in wk 2 postpartum for cows fed HF.