title: Cigarette smoke exposure impairs VEGF-induced endothelial cell migration: role of NO and reactive oxygen species.
abstract: Endothelial dysfunction is one of the earliest pathological effects of cigarette smoking. Vascular endothelial growth factor (VEGF) has been shown to be an important regulator of endothelial healing and growth. Accordingly, we tested the hypothesis that cigarette smoke exposure impairs VEGF actions in endothelial cells. In human umbilical vein endothelial cells (HUVECs), cigarette smoke extracts (CSE) inhibited VEGF-induced tube formation in the matrigel assay. CSE did not affect HUVECs proliferation, but significantly reduced cellular migration in response to VEGF. This impaired migratory activity was associated with a reduced expression of alpha(v)beta(3), alpha(v)beta(5), alpha(5)beta(1) and alpha(2)beta(1) integrins. The Akt/eNOS/NO pathway has been shown to be important for VEGF-induced endothelial cell migration. We found that CSE inhibited Akt/eNOS phosphorylation and NO release in VEGF-stimulated HUVECs. This was associated with an increased generation of reactive oxygen species (ROS). Importantly, in HUVECs exposed to CSE, treatment with antioxidants (NAC, vitamin C) reduced ROS formation and rescued VEGF-induced NO release, cellular migration and tube formation. Moreover, treatment with NO donors (SNAP, SNP) or a cGMP analog (8-Br-cGMP) rescued integrin expression, cellular migration and tube formation in endothelial cells exposed to CSE. (1) Cigarette smoke exposure impairs VEGF-induced endothelial cell migration and tube formation. (2) The mechanism involves increased generation of ROS, decreased expression of surface integrins together with a blockade of the Akt/eNOS/NO pathway. (3) These findings could contribute to explain the negative effect of cigarette smoking on endothelial function and vessel growth.
authors.affiliation: Department of Cardiovascular Research, Centre Hospitalier de l'Université de Montréal, Québec, Canada.
authors.forename: Sophie Elise
authors.initials: SE
authors.lastname: Michaud
authors.forename: Sylvie
authors.initials: S
authors.lastname: Dussault
authors.forename: Jessika
authors.initials: J
authors.lastname: Groleau
authors.forename: Paola
authors.initials: P
authors.lastname: Haddad
authors.forename: Alain
authors.initials: A
authors.lastname: Rivard
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D002318:Cardiovascular Diseases / Q000209:etiology; D002465:Cell Movement / Q000187:drug effects*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D004347:Drug Interactions; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D016023:Integrins / Q000096:biosynthesis; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012307:Risk Factors; D026423:S-Nitroso-N-Acetylpenicillamine / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D012906:Smoke / Q000009:adverse effects*; D012907:Smoking / Q000009:adverse effects*; D013873:Thionucleotides / Q000494:pharmacology; D014026:Nicotiana / Q000009:adverse effects*; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D016023:Integrins; D020030:Nitric Oxide Donors; D017382:Reactive Oxygen Species; D012906:Smoke; D013873:Thionucleotides; D042461:Vascular Endothelial Growth Factor A; C083763:8-bromoguanosino-3',5'-cyclic monophosphorothioate; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D006152:Cyclic GMP