The mechanism by which homocysteine (Hcy) causes endothelial dysfunction is probably mediated by oxidative stress. The aim of this study was to evaluate the effect of oxidative stress on endothelial function in young and elderly hyperhomocysteinemic (HHcy) men. A total of 35 HHcy (Hcy > 15 micro mol/L), young (n = 15; 20-40 y) and elderly men (n = 20; > 65 y) and 33 normohomocysteinemic (NHcy; controls) young (n = 14) and elderly (n = 19) men (Hcy < 13 micro mol/L), without classic cardiovascular risk factors were recruited. Serum Hcy, folate, and vitamin B-12, whole-blood glutathione, plasma total antioxidants status, TBARS, and 8-F(2alpha) isoprostanes were determined. Noninvasive ultrasound measurements of endothelium-dependent (EDVR) and -independent dilatation (EIVR) were performed. EDVR, EIVR, and markers of oxidative stress did not differ among the groups. Folate concentrations were higher in elderly than in young men (P < 0.001), independent of Hcy concentrations. Vitamin B-12 concentrations were lower in HHcy than in NHcy elderly men (P < 0.045). EDVR was correlated with folate concentrations in young men (r = 0.40, P = 0.04) and negatively with BMI in elderly men (r = -0.52, P = 0.002). In the present study, HHcy with normal serum folate concentrations was not associated with poor EDVR or oxidative stress in healthy young and elderly men.