The present study aimed to investigate the molecular mechanisms of the ameliorative effects of chronic aerobic exercise on non‑alcoholic steatohepatitis (NASH) in rat skeletal muscle. Female Sprague‑Dawley rats (n=6‑9 per group) were divided into four groups: i) Rats fed with normal chow; ii) exercise rats fed with normal chow + exercise (run on a rotarod for 30 min per day from 9‑12 weeks); iii) rats fed with a high‑fat diet (HFD); iv) rats fed with an HFD + exercise. All HFD rats were fed with an HFD consisting of 30% fat from fish oil throughout the study for 12 weeks. Exercise decreased the levels of hepatic lipogenic markers carbohydrate‑responsive element‑binding protein, fat‑specific protein 27 and liver X receptor and improved systemic glucose and insulin intolerance in the NASH animal model. The beneficial effects may have been mediated partly via the tripartite motif‑containing family protein 72 (TRIM72)/PI3K/Akt/mTOR pathway, accompanied with an upregulation of glucose transporter 4 in the skeletal muscle. The exercise regimen activated the master regulator of antioxidant enzymes, nuclear factor erythroid 2‑related factor 2, with upregulation of superoxide dismutase [Cu‑Zn] expression and a corresponding decrease in kelch‑like ECH‑associated protein 1 expression, but failed to decrease the levels of the oxidative marker malondialdehyde in the HFD rat skeletal muscle. Chronic exercise decreased the expression of the inflammation marker NF‑κB, followed by a decrease in interleukin‑6 and tumor necrosis factor‑α levels, as verified by a corresponding increase in the level of NF‑κB inhibitor α expression. Exercise may exert its beneficial effects by improving muscle insulin sensitivity via the TRIM72/PI3K/Akt/mTOR pathway, contributing to the improvement of systemic insulin intolerance, and finally leading to decreased hepatic lipogenesis during NASH. The attenuation of insulin resistance by exercise may be partly achieved through a decrease in the level of inflammation and an increased antioxidant response.