Physical activity affects the vasculature through mechanisms related to nitric oxide bioavailability, oxidative stress, and inflammation; with endothelial function at the centre of this triad. In a South African setting, with the prevalence of hypertension and physical inactivity being alarmingly high, we aimed to investigate relationships of vascular function with markers of oxidative stress, inflammation and nitric oxide synthesis capacity in physically active and inactive groups. Based on the 2010 World Health Organisation guidelines, black and white school teachers were divided into physically active (n = 84) and physically inactive (n = 132) groups. Twenty-four-hour blood pressure (24 h BP), total peripheral resistance and Windkessel compliance were measured. Markers of oxidative stress, inflammation and nitric oxide synthesis capacity were analysed. Windkessel compliance (p = 0.041) and homoarginine (p = 0.006) were higher in the physically active group. In the same group, 24 h diastolic BP associated with total glutathione (β = 0.17; p = 0.056), and 24 h BP (systolic blood pressure: β = 0.23, p = 0.006; diastolic blood pressure: β = 0.22, p = 0.019) associated with homoarginine. In the physically inactive group, 24 h BP (systolic blood pressure: β = 0.26, p < 0.001; diastolic blood pressure: β = 0.23, p = 0.007) associated with symmetric dimethylarginine (SDMA). These associations were independent of inflammation. Despite only reaching moderate physical activity levels, vascular function and nitric oxide synthesis capacity were more favourable in the physically active population compared to the physically inactive population. These results may suggest that even moderate physical activity could increase nitric oxide synthesis capacity, which in turn may mitigate the development of cardiovascular disease in this population.