Endothelial cell (EC) migration plays an important role in wound repair of blood vessels. We have previously reported that eicosapentaenoic acid (EPA; 20:5, n-3) pretreatment stimulates migration of ECs but not smooth muscle cells. In the present study, we used the modified Boyden chamber technique to investigate whether the stimulative effect of EPA pretreatment on EC migration is caused by EPA itself or by some metabolites of EPA. When ECs were treated with EPA (5 micrograms/ml) for 2 days, EPA was predominantly elongated to docosapentaenoic acid (DPA; 22:5, n-3), with little docosahexaenoic acid (DHA; 22:6, n-3) being formed. Direct pretreatment of ECs with DPA (0.01-1.0 microgram/ml) resulted in a dose-dependent increase in migration in response to fetal bovine serum. Moreover, maximum stimulation of EC migration by DPA pretreatment (0.5 microgram/ml) was achieved at a concentration one-tenth of that required for maximal stimulation by EPA pretreatment (5.0 micrograms/ml), indicating that DPA is a potent stimulator of EC migration. We have demonstrated by lipid analysis that direct DPA pretreatment (0.5 microgram/ml) sufficiently increased the absolute quantity of phospholipids of ECs. Cyclooxygenase inhibitor and lipoxygenase inhibitor did not abolish the stimulative effect of DPA pretreatment on EC migration. In contrast to EC migration, DPA pretreatment had no effect on smooth muscle cell migration. Together these data suggest that the stimulative effect of EPA on EC migration occurs via DPA, and that DPA may act as a powerful anti-atherogenic factor.