Atherosclerosis is a systemic disease with focal cardiovascular events. Although the accelerated development of peripheral arterial disease in diabetic patients is well known, the pathogenic mechanism of site-specific susceptibility to glycemia is uncertain. To investigate the association of fasting and post-challenge glucose levels with intima-media thickness (IMT) at different arterial sites. Forty consecutive middle-aged volunteers aged 37 to 53 years were recruited to define the association of IMT with cardiovascular risk factors at 12 carotid and 6 lower-limb arterial sites. A linear mixed model was used to regress the primary outcome measures, which were repeated measures of IMT at multiple arterial sites (18 sites per participant), on fixed-effect predictors of various conventional cardiovascular risk factors, while accounting for the interdependence of repeated measures taken from the same participant with unstructured covariance. Carotid IMTs were associated independently with waist circumference and systolic blood pressure, whereas lower-limb IMTs were associated with waist circumference, low-density lipoprotein cholesterol (LDL-C), glycated hemoglobin A1C (HbA1C), and fasting and 2-hour post-challenge plasma glucose levels; these associations were stronger in overall arteries. Independent associations of ALT and smoking with IMT appeared only in overall arteries. In a middle-aged, nonclinical sample, lower-limb but not carotid IMTs are associated independently with HbA1C, and fasting and 2-hour post-challenge plasma glucose levels.