Objective To study the protective effect of nobiletin on hepatocyte apoptosis in rat model of nonalcoholic fatty liver disease (NAFLD). Methods 72 SD rats were randomly divided into 6 groups: the control group, model group, positive drug silymarin treatment group (14 mg/kg), and nobiletin low dose (100 mg/kg), medium dose (200 mg/kg) and high dose (400 mg/kg) treatment groups. NAFLD rat model was established by high-fat diet for 8 weeks followed by 6 weeks of treatment. After scarification the liver pathology was evaluated by HE staining, liver index was calculated, and serum levels of ALT, AST, TC, TG, LDL-C, HDL-C, IL-1a, IL-1b and IL-6 were measured. Protein expression and mRNA levels of TNF-α, TNFR-1, Fas, FasL, caspase-8, caspase-3, Bcl-2 and Bax in liver tissue were detected by Western Blot and qRT-PCR. Results Compared with the control group, rats in model group were apathetic, and rats in the silymarin group, low dose, medium dose and high dose nobiletin groups were obviously alleviated. In the model group, increase of the liver wet weight, liver index, ALT, AST, TG, TC, LDL-C, IL-1a, IL-1b and IL-6, also decrease of HDL-C(P<0.05) was observed. Compared with the model group, the liver wet weight, liver index, ALT, AST, TG, TC, LDL-C, IL-1a, IL-1b and IL-6 in the silymarin group and the low dose, medium dose and high dose nobiletin groups were significantly decreased (P<0.05) along with significant increase of HDL-C(P<0.05). In addition, the liver index, ALT, AST, IL-1a, IL-1b and IL-6 in the high dose nobiletin group were significantly lower than that of the silymarin group(P<0.05), and the TG, TC and LDL-C in the middle dose and high dose nobiletin groups were significantly lower than that of the silymarin group(P<0.05), while HDL-C in the middle dose and high dose nobiletin groups were significantly higher than that of the silymarin group(P<0.05). There were no significant difference in the body weight. In the model group, TNF-α, TNFR-1, FasL, Fas, Bax, caspase-8 and caspase-3 protein and mRNA levels were significantly increased(P<0.05), and Bcl-2 protein and mRNA levels were significantly declined (P<0.05). Compared with the model group, the TNF-α, TNFR-1, FasL, Fas, caspase-8 and caspase-3 protein and mRNA levels were significantly decreased in the silymarin and the low dose, medium dose and high dose nobiletin groups(P<0.05), and the expression of Bcl-2 in the silymarin treatment group and the low dose, medium dose and high dose nobiletin groups were significantly increased(P<0.05). Conclusion The nobiletin has obvious protective effect on the NAFLD rats induced by high-fat diet, and the mechanism of action was attribute to the inhibition of nobiletin in apoptosis-related proteins expression and inflammatory response.