Arterial stiffness has been shown to be a subclinical marker associated with cardiovascular disease. Meanwhile, long-term exercise has been demonstrated to reduce arterial stiffness, providing a decrease in cardiovascular risk. However, the acute effect of exercise on arterial stiffness is unclear. This systematic review and meta-analysis aimed to assess the acute effect of exercise interventions on arterial stiffness in healthy adults. We searched the Cochrane Central Register of Controlled Trials, MEDLINE (via Pubmed), Scopus, and Web of Science databases, from their inception to 30 June 2020. A meta-analysis was performed to evaluate the acute effect of exercise on arterial stiffness using random-effects models to calculate pooled effect size estimates and their corresponding 95% CI. Pulse wave velocity was measured as an arterial stiffness index. The 30 studies included in the meta-analysis showed that pulse wave velocity was not modified immediately after exercise (0 min post) (ES: 0.02; 95% CI: −0.22, 0.26), but subsequently decreased 30 min after exercise (ES: −0.27; 95% CI: −0.43, −0.12). Thereafter, pulse wave velocity increased to its initial value 24 h after exercise (ES: −0.07; 95% CI: −0.21, 0.07). Our results show that, although there is a significant reduction in pulse wave velocity 30 min after exercise, the levels of arterial stiffness return to their basal levels after 24 h. These findings could imply that, in order to achieve improvements in pulse wave velocity, exercise should be performed on a daily basis. Keywords: arterial stiffness, pulse wave velocity, exercise, acute effect, healthy adults