Background: Non-alcoholic fatty liver disease (NAFLD) is a frequent progressive disorder manifested by fat accumulation in the liver and usually related to obesity and insulin resistance, but its pathogenesis is still uncertain. Nesfatin-1 is a polypeptide derived from nucleobindin-2 and involved in regulation of food intake and glucose homeostasis. The relationship between nesfatin-1 and NAFLD is still controversial. Objective: To evaluate serum levels of nesfatin-1 in NAFLD model induced by high fat diet (HFD) in male albino rats. Material and methods: Forty eight male adult albino rats were divided into four equal groups: 2 control groups that were fed ordinary diet for 4 weeks (group IA) and 12 weeks (group IB), and 2 HFD groups that were fed HFD for 4 weeks (group IIA) and 12 weeks (group IIB). In all groups, abdominal circumference, body weight, serum levels of nesfatin-1, insulin, glucose, C-reactive protein (CRP), lipid profile parameters, and liver enzymes (ALT & AST) were measured. BMI and HOMA-IR were calculated, and isolated liver tissues were examined histopathologically. Results: After 4-week and 12-week-HFD feeding, the rats developed simple steatosis and steatohepatitis, respectively. These were proved by the progressive rise of liver enzymes and the histopathological findings. Besides, there was a significant progressive rise in BMI, HOMA-IR, serum levels of nesfatin-1, glucose, insulin, CRP, and all lipid profile parameters except high density lipoprotein that significantly decreased in HFD groups in comparison to control groups. Moreover, nesfatin-1 correlated positively with all measured parameters in HFD groups except for HDL that showed negative correlation with nesfatin-1. Conclusion: Serum levels of nesfatin-1 increased in NAFLD rat model induced by HFD. This rise may be attributed to feeding rats with HFD, hyperglycemia or may compensate for the inflammation and disturbed metabolism.