Vascular aging is characterized by increased oxidative stress and pro-inflammatory phenotypic alterations. Metabolic stress, such as hyperglycemia in diabetes, is known to increase production of reactive oxygen species (ROS) and promote inflammatory gene expression accelerating vascular aging. The oxidative stress hypothesis of aging predicts that vascular cells of long-lived species exhibit lower steady-state production of ROS and/or superior resistance to the pro-oxidant effects of metabolic stress. We tested this hypothesis using two taxonomically related rodents, the white-footed mouse (Peromyscus leucopus) and the house mouse (Mus musculus) that show a more than two-fold difference in maximum lifespan potential (8.2 and 3.5 years, respectively). We compared interspecies differences in steady-state and high glucose (HG; 30 mmol/L)-induced production of O₂^.- and H₂O₂, endothelial function, mitochondrial ROS generation and inflammatory gene expression in cultured aortic segments. In P.leucopus aortas steady-state endothelial O₂^.- and H₂O₂ production and ROS generation by mitochondria were less than in M.musculus vessels. Furthermore, vessels of P.leucopus were more resistant to the pro-oxidant effects of HG. Primary fibroblasts from P.leucopus also exhibited less steady-state and HG-induced ROS production than M.musculus cells. In M.musculus arteries HG elicited significant up-regulation of inflammatory markers (TNF, IL-6, ICAM-1, VCAM and MCP-1). In contrast, the pro-inflammatory effects of HG were blunted in P.leucopus vessels. Thus, increased lifespan potential in P.leucopus is associated with a decreased cellular ROS generation and increased resistance to the pro-oxidant and pro-inflammatory effects of metabolic stress, which accord with the predictions of the oxidative stress hypothesis of aging.