Endothelial dysfunction and inflammation are characteristics of subclinical atherosclerosis and may increase through progressive menopausal stages. Evaluating endothelial responses to acute exercise can reveal underlying dysfunction not apparent in resting conditions. The purpose of this study was to investigate markers of endothelial function and inflammation before and after acute exercise in healthy low active perimenopausal (PERI) and late postmenopausal (POST) women. Flow mediated dilation (FMD), CD31⁺/CD42b^- and CD62E⁺ endothelial microparticles (EMPs), and the circulating inflammatory factors monocyte chemoattractant protein 1 (MCP-1), interleukin 8 (IL-8), and tumor necrosis factor alpha (TNFα), were measured before and 30 minutes following acute exercise. Before exercise, FMD was not different between groups (PERI:6.4±0.9% vs. POST:6.5±0.8 %, p=0.97); however, following acute exercise PERI tended to improve FMD (8.5±0.9%, p=0.09) whereas POST did not (6.2±0.8%, p=0.77). Independent of exercise, we observed transient endothelial dysfunction in POST with repeated FMD measures. There was a group x exercise interaction for CD31⁺/CD42b^- EMPs (p=0.04), where CD31⁺/CD42b^- EMPs were similar before exercise (PERI:57.0±6.7 EMPs/μl vs. POST:58.5±5.3 EMPs/μl, p=0.86) but were higher in POST following exercise (PERI:48.2±6.7 EMPs/μl vs. POST:69.4±5.3 EMPs/μl, p=0.023). CD62E⁺ EMPs were lower in PERI compared with POST before exercise (p<0.001) and increased in PERI (p=0.04) but did not change in POST (p=0.68) in response to acute exercise. Following acute exercise, MCP-1 (p=0.055), TNFα (p= 0.02), and IL-8 (p<0.001) were lower in PERI but only IL-8 decreased in POST (p<0.001). Overall, these data suggest that perimenopausal and late postmenopausal women display different endothelial and inflammatory responses to acute exercise.