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To investigate the relationship between 1-h post-load plasma glucose, measured during an oral glucose tolerance test, and arterial stiffness, determined by brachial–ankle pulse-wave velocity, in normotensive subjects with normal glucose tolerance.Study subjects were non-industrial workers aged 25–55 years (n = 8381) who underwent a regular health check-up every 5 years. We included only normotensive subjects with normal glucose tolerance based on the American Diabetes Association criteria. Subjects taking medication and having an abnormal ankle–brachial index (≤1.0 or ≥1.3) were excluded. The final sample comprised 4970 participants (mean age: 38.8 ± 9.4 years; women: n = 2048).1-h post-load plasma glucose correlated with brachial–ankle pulse-wave velocity in men (β = 0.04, p = 0.01), but not women (β = –0.03, p = 0.13) in multivariate linear regression analysis. We found a significant interaction between 1-h post-load plasma glucose and age in men (p = 0.04); therefore, a subgroup analysis was performed in each 5-year age group. The correlation between 1-h post-load plasma glucose and brachial–ankle pulse-wave velocity was significant in the 55-year-old age group (β = 0.12, p = 0.01) and neared significant in 45-year-old (β = 0.08, p = 0.07) and 50-year-old (β = 0.09, p = 0.07) age groups.Elevated 1-h post-load plasma glucose levels were associated with arterial stiffness in normotensive, middle-aged men with normal glucose tolerance.