Non-alcoholic fatty liver disease (NAFLD) is associated with hepatic insulin resistance, but the exact mechanism remains unclear. This study investigated the relationship between hepatic fat accumulation and hepatic insulin resistance in rats. Short-term high-fat feeding caused significant hepatic fat accumulation without significant peripheral fat accumulation. This model allowed researchers to study the effects of hepatic fat on insulin resistance without confounding factors from peripheral fat. The study found that hepatic insulin resistance was linked to impaired insulin signaling, specifically reduced tyrosine phosphorylation of IRS-1 and IRS-2, and activation of PKC-ε and JNK1. These changes were associated with decreased insulin activation of glycogen synthase and increased gluconeogenesis. Treatment with 2,4-dinitrophenol, which increases energy expenditure, prevented hepatic fat accumulation and improved insulin sensitivity by inhibiting PKC-ε and JNK1 activation. The results suggest that hepatic fat accumulation leads to insulin resistance by stimulating gluconeogenesis and activating PKC-ε and JNK1, which interfere with insulin signaling. The study supports the hypothesis that hepatic fat accumulation is a key factor in the development of hepatic insulin resistance.Non-alcoholic fatty liver disease (NAFLD) is associated with hepatic insulin resistance, but the exact mechanism remains unclear. This study investigated the relationship between hepatic fat accumulation and hepatic insulin resistance in rats. Short-term high-fat feeding caused significant hepatic fat accumulation without significant peripheral fat accumulation. This model allowed researchers to study the effects of hepatic fat on insulin resistance without confounding factors from peripheral fat. The study found that hepatic insulin resistance was linked to impaired insulin signaling, specifically reduced tyrosine phosphorylation of IRS-1 and IRS-2, and activation of PKC-ε and JNK1. These changes were associated with decreased insulin activation of glycogen synthase and increased gluconeogenesis. Treatment with 2,4-dinitrophenol, which increases energy expenditure, prevented hepatic fat accumulation and improved insulin sensitivity by inhibiting PKC-ε and JNK1 activation. The results suggest that hepatic fat accumulation leads to insulin resistance by stimulating gluconeogenesis and activating PKC-ε and JNK1, which interfere with insulin signaling. The study supports the hypothesis that hepatic fat accumulation is a key factor in the development of hepatic insulin resistance.