Purpose The purpose of this study was to investigate the effect of treadmill exercise and MitoQ treatment on NADPH oxidase, antioxidation and vascular function-related factors in aortic of D-galactose-induced aging rats. Methods To induce the animal model of aging, D-galactose was diluted in saline, and a dose of 100mg /kg was intraperitoneally injected into Sprague-Dawley rats once a day for a total of 10 weeks. Rats were divided into five groups: Control group (CON, n=9), D-galactose control group (DC, n=9), D-galactose+MitoQ group (DM, n=9), D-galactose+Exercise group (DE, n=9), D-galactose+MitoQ + Exercise group(DME, n=9), and treadmill exercise was conducted for 5 days/week during 8 weeks with gradual increase of intensity. MitoQ treatment was intraperitoneally injected at a concentration of 100μM/kg twice a week for 8 weeks during the research period. Results The result showed that treadmill exercise and MitoQ treatment decreased the expression of NADPH oxidase level and increased antioxidant enzyme such as SOD-2, catalase. It lead to positive effects such as increasing the level of eNOS, a protein involved in vascular function while decreasing the level of VCAM-1. In addition, as a result, it showed the structurally reduced intima-media thickness. Conclusions It can be concluded that treadmill exercise and MitoQ treatment are effective in ameliorating and treating vascular dysfunction resulting from aging.