The TT genotype of C677T polymorphism in 5,10-methylenetetrahydrofolate reductase (MTHFR) induces elevation of homocysteine level and leads to atherosclerosis and arterial stiffening. Furthermore, cardiorespiratory fitness level is also associated with arterial stiffness. In the present study, a cross-sectional investigation of 763 Japanese men and women (18 -70 yr old) was performed to clarify the effects of cardiorespiratory fitness on the relationship between arterial stiffness and MTHFR C677T gene polymorphism. Arterial stiffness was assessed by carotid -stiffness with ultrasonography and tonometry. The study subjects were divided into high-cardiorespiratory fitness (HighFit) and low-cardiorespiratory fitness (Low-Fit) groups based on the median value of peak oxygen uptake in each sex and decade. The plasma homocysteine level was higher in the TT genotype of MTHFR C677T polymorphism compared with CC and CT genotype individuals. MTHFR C677T polymorphism showed no effect on carotid -stiffness, but there was a significant interaction effect between fitness and MTHFR C677T polymorphism on carotid -stiffness (P ϭ 0.0017). In the Low-Fit subjects, carotid -stiffness was significantly higher in individuals with the TT genotype than the CC and CT genotypes. However, there were no such differences in High-Fit subjects. In addition, -stiffness and plasma homocysteine levels were positively correlated in Low-Fit subjects with the TT genotype (r ϭ 0.71, P Ͻ 0.0001), but no such correlations were observed in High-Fit subjects. In CC and CT genotype individuals, there were also no such correlations in either fitness level. These results suggest that the higher cardiorespiratory fitness may attenuate central artery stiffening associated with MTHFR C677T polymorphism. peak oxygen uptake; arterial stiffness; homocysteine; 5,10-methylenetetrahydrofolate reductase ELEVATED PLASMA HOMOCYSTEINE level is considered a risk factor for cardiovascular events and is associated with arterial stiffness and atherosclerosis in subjects with some cardiovascular risk factors (7,15,30,36). High homocysteine levels may impair endothelial function, increase oxidative stress, and alter protein structure (5, 6, 37). Exposure of endothelial cells to elevated homocysteine levels results in decreased availability of nitric oxide (NO), which has vasodilatory and antiplatelet effects, and impaired vascular function, which are early events in atherogenesis (6,29,33,35). Homocysteine metabolism represents an interesting model of gene-environment interaction (34,38). Elevations in homocysteine may be caused by genetic and environmental factors and by gene-gene and/or geneenvironment interactions. The enzyme 5,10-methylenetetrahydrofolate reductase (MTHFR) catalyzes the irreversible conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate (4). A polymorphism of C677T (Ala¡Val) in the gene encoding MTHFR is associated with decreased activity of the enzyme due to thermolability (1). In individuals homozygous for the T (Val) allele, a r...