The impact of walking on cardiovascular health, as compared with vigorous physical activity, remains controversial. We performed a cross-sectional analysis including 185 healthy participants drawn from the Whitehall II epidemiological cohort to examine whether there is an independent association between walking and markers of hemostasis and inflammation after controlling for vigorous physical activity. Blood was drawn for the assessment of von Willebrand factor antigen (vWF), fibrinogen, interleukin (IL)-6, IL-1 receptor antagonist (IL-1ra), and tumor necrosis factor-α (TNF-α). Self-reported levels and types of physical activity were recorded, including time spent walking per week and frequency of participation in vigorous physical activity. Multiple linear regression analyses, adjusted for age, body mass index, blood pressure, gender, smoking, alcohol, grade of employment, and frequency of vigorous physical activity revealed that time spent walking was inversely related to vWF (β=−0.13, P=0.086), fibrinogen (β=−0.16, P=0.016), IL-6 (β=−0.12, P=0.087), and TNF-α (β=−0.16, P=0.039). In addition, vigorous activity was inversely associated with hemostatic markers but not with inflammatory cytokines. Regular walking is associated with lower levels of hemostatic and inflammatory markers independently of vigorous physical activity in healthy men and women.