title: Cigarette smoke causes lung vascular barrier dysfunction via oxidative stress-mediated inhibition of RhoA and focal adhesion kinase.
abstract: Cigarette smoke (CS) is a major cause of chronic lung and cardiovascular diseases. Recent studies indicate that tobacco use is also a risk factor for acute lung injury (ALI) associated with blunt trauma. Increased endothelial cell (EC) permeability is a hallmark of ALI. CS increases EC permeability in vitro and in vivo; however, the underlying mechanism is not well understood. In this study, we found that only 6 h of exposure to CS impaired endothelial barrier function in vivo, an effect associated with increased oxidative stress in the lungs and attenuated by the antioxidant N-acetylcysteine (NAC). CS also exacerbated lipopolysaccharide (LPS)-induced increase in vascular permeability in vivo. Similar additive effects were also seen in cultured lung EC exposed to cigarette smoke extract (CSE) and LPS. We further demonstrated that CSE caused disruption of focal adhesion complexes (FAC), F-actin fibers, and adherens junctions (AJ) and decreased activities of RhoA and focal adhesion kinase (FAK) in cultured lung EC. CSE-induced inhibition of RhoA and FAK, endothelial barrier dysfunction, and disassembly of FAC, F-actin, and AJ were prevented by NAC. In addition, the deleterious effects of CSE on FAC, F-actin fibers, and AJ were blunted by overexpression of constitutively active RhoA and of FAK. Our data indicate that CS causes endothelial barrier dysfunction via oxidative stress-mediated inhibition of RhoA and FAK.
authors: Vascular Research Laboratory, Providence Veterans Affairs Medical Center, Department of Medicine, Alpert Medical School of Brown University, RI, USA. Qing_Lu@brown.edu
authors: Qing
authors: Q
authors: Lu
authors: Pavlo
authors: P
authors: Sakhatskyy
authors: Katie
authors: K
authors: Grinnell
authors: Julie
authors: J
authors: Newton
authors: Melanie
authors: M
authors: Ortiz
authors: Yulian
authors: Y
authors: Wang
authors: Juan
authors: J
authors: Sanchez-Esteban
authors: Elizabeth O
authors: EO
authors: Harrington
authors: Sharon
authors: S
authors: Rounds
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000199:Actins / Q000378:metabolism; D022005:Adherens Junctions / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D017097:Electric Impedance; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D022001:Focal Adhesions / Q000378:metabolism; D008070:Lipopolysaccharides; D008168:Lung / Q000098:blood supply / Q000187:drug effects / Q000473:pathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018384:Oxidative Stress; D010539:Permeability / Q000187:drug effects; D061251:Primary Cell Culture; D011654:Pulmonary Edema / Q000139:chemically induced; D012906:Smoke / Q000009:adverse effects*; D012907:Smoking / Q000009:adverse effects*; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D000199:Actins; D000975:Antioxidants; D008070:Lipopolysaccharides; D012906:Smoke; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496051:Ptk2 protein, mouse; C526380:RhoA protein, mouse; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; D000111:Acetylcysteine