Endothelial nitric oxide synthase inhibition does not alter endothelial progenitor cell colony forming capacity or migratory activity

Abstract

Endothelial nitric oxide synthase (eNOS) activity has been shown to play a pivotal role in the mobilization of endothelial progenitor cells (EPCs) into the circulation from bone marrow. Indeed, in eNOS-deficient mice, exercise-induced EPC mobilization is severely diminished. We determined ex vivo whether circulating EPC colony-forming capacity and migratory activity are influenced by eNOS activity. Peripheral-blood mononuclear cells were isolated from 20 healthy adults and preplated for 2 days, and nonadherent cells were further cultured for 7 days in the presence and absence of N-nitro-L-arginine methyl ester (L-NAME, 300 microM, an eNOS antagonist) to determine EPC colony-forming units. Migratory activity of EPCs, cultured with and without L-NAME (300 microM) was determined by utilizing a modified Boyden chamber. The number of EPC colony-forming units was not significantly different when cultured in the absence or presence of L-NAME (21+/-5 vs 18+/-5). Moreover, eNOS inhibition did not alter EPC migratory activity; mean fluorescence was similar in samples cultured with (983+/-126 RFUs) and without (962+/-105 RFUs) L-NAME. These in vitro results suggest that, in contrast to EPC mobilization from the bone marrow, eNOS does not exert a modulatory influence on the functional capacity of circulating EPCs to either form colonies or migrate.