Elevated circulating alanine aminotransferase (ALT) and γ-glutamyltransferase (γ-GTP) activities, and the accumulation of fat, particularly visceral fat, in healthy and preclinical subjects reportedly increase the risk for metabolic diseases such as diabetes. In the present study, we examined the associations between these hepatic enzymes and the total visceral and subcutaneous fat area, and for both regions of fat independently, in healthy and preclinical middle-aged Japanese men. We conducted a cross-sectional study of men who participated in health check-ups in Japan. We removed participants, who were diagnosed with metabolic diseases at the time of the health check-up. Three hundred fifteen subjects aged 40-64 y (mean±SD, 50.5±6.9 y) were selected. We compared associations between the total visceral and subcutaneous fat area, and for both regions independently, with various clinical parameters, including hepatic enzyme markers, using Spearman's rank correlation coefficient analysis and multiple linear regression analysis. The total visceral and subcutaneous fat area and both regions independently were positively associated with body mass index, systolic and diastolic blood pressure, fasting blood glucose, total cholesterol, low-density lipoprotein cholesterol, aspartate aminotransferase, ALT and γ-GTP. ALT and γ-GTP activities were the strongest explanatory variables for increased visceral fat area, independent of the subcutaneous fat area. In contrast, these hepatic enzymes were not explanatory variables for increased subcutaneous fat area. The results of the present study show that the accumulation of visceral fat is positively associated with ALT and γ-GTP activities independently of subcutaneous fat area in healthy and preclinical Japanese men.