Background and Objective: The Western diet, characterized by a high content of fats and simple carbohydrates, is considered one of the main contributors to hepatic metabolic disorders, including increased lipogenesis and hepatic steatosis. In this context, non-pharmacological interventions such as exercise training and novel nutritional strategies, including ketone ester supplementation, have gained attention as potential approaches for modulating lipogenic pathways. The aim of the present study was to investigate the effects of high-intensity interval training (HIIT) and ketone ester supplementation, alone and in combination, on hepatic lipogenic protein markers in Western diet–fed mice.

Methods: This experimental, randomized, and controlled study was conducted on 30 male C57BL/6J mice. After an acclimatization period, the animals were randomly assigned to five groups: normal diet (ND), Western diet (WD), Western diet plus ketone ester supplementation (WD+S), Western diet plus HIIT (WD+EX), and Western diet plus combined HIIT and ketone ester supplementation (WD+EX+S). The HIIT protocol was performed for 4 weeks, with 3 sessions per week, on a treadmill set at a 5° incline. Each training session consisted of 10 bouts of 2-minute exercise intervals at 85–90% of maximal running speed, interspersed with 2-minute active recovery periods at 50% of maximal running speed. Exercise speed was progressively increased from 19 m/min in the first week to 22 m/min in the fourth week. Ketone ester supplementation was administered daily via oral gavage using a dedicated gavage syringe. The dose was set at 5 mg per kilogram of body weight and was adjusted weekly based on the body weight of each rat. At the end of the intervention period, hepatic protein levels of sterol regulatory element–binding protein-1c (SREBP-1c), carbohydrate-responsive element–binding protein (ChREBP), and peroxisome proliferator-activated receptor gamma (PPAR-γ) were measured using Western blot analysis. Data were analyzed using independent t-tests, one-way analysis of variance (ANOVA), and Tukey’s post hoc test.

Results: The results demonstrated that consumption of the Western diet significantly increased hepatic protein levels of SREBP-1c, ChREBP, and PPAR-γ (P