Nonalcoholic fatty liver disease (NAFLD) currently represents the most common hepatic disease worldwide and is closely linked to cardiovascular disease, obesity and diabetes mellitus. This study aimed to investigate NAFLD and its influence on different monocyte subpopulations to determine the presence of significant associations. A total of 3 monocyte subpopulations were investigated, i.e. classical (CD14++CD16−), intermediate (CD14++CD16+) and non-classical (CD14+CD16++). Of the participants 261 were included in this study (n = 53 with NAFLD, n = 208 controls). Ultrasonography was used to diagnose NAFLD and exclude other morphologic causes of liver diseases and other tests (including medical history inquiries and detection of hepatitis virus) were performed to exclude other causes of parenchymal liver disease. Classical inflammatory and metabolic-related NAFLD biomarkers were also determined. In contrast to the healthy control group, the intermediate monocyte fraction was increased in NAFLD patients (p = 0.032), while the classical monocyte fraction was decreased (p = 0.025). Intermediate monocyte fraction, body mass index (BMI) and tumor necrosis factor alpha (TNF-α) were independent risk factors for NAFLD. Classical, non-classical and intermediate monocytes fraction were strongly associated with age, triglyceride, and waist circumference. This study suggests that the intermediate monocyte fraction in peripheral blood is likely related to the aggravation of NAFLD.