title: Involvement of ROCK-mediated endothelial tension development in neutrophil-stimulated microvascular leakage.
abstract: Neutrophil-induced coronary microvascular barrier dysfunction is an important pathophysiological event in heart disease. Currently, the precise cellular and molecular mechanisms of neutrophil-induced microvascular leakage are not clear. The aim of this study was to test the hypothesis that rho kinase (ROCK) increases coronary venular permeability in association with elevated endothelial tension. We assessed permeability to albumin (P(a)) in isolated porcine coronary venules and in coronary venular endothelial cell (CVEC) monolayers. Endothelial barrier function was also evaluated by measuring transendothelial electrical resistance (TER) of CVEC monolayers. In parallel, we measured isometric tension of CVECs grown on collagen gels. Transference of constitutively active (ca)-ROCK protein into isolated coronary venules or CVEC monolayers caused a significant increase in P(a) and decreased TER in CVECs. The ROCK inhibitor Y-27632 blocked the ca-ROCK-induced changes. C5a-activated neutrophils (10(6)/ml) also significantly elevated venular P(a), which was dose-dependently inhibited by Y-27632 and a structurally distinct ROCK inhibitor, H-1152. In CVEC monolayers, activated neutrophils increased permeability with a concomitant elevation in isometric tension, both of which were inhibited by Y-27632 or H-1152. Treatment with ca-ROCK also significantly increased CVEC monolayer permeability and isometric tension, coupled with actin polymerization and elevated phosphorylation of myosin regulatory light chain on Thr18/Ser19. The data suggest that during neutrophil activation, ROCK promotes microvascular leakage in association with actin-myosin-mediated tension development in endothelial cells.
authors.affiliation: UCDMC Dept. of Surgery, Div. of Research, 2805 50th St., Rm. 2411, Sacramento, CA 95817, USA. jwbreslin@ucdavis.edu
authors.forename: Jerome W
authors.initials: JW
authors.lastname: Breslin
authors.forename: Hengrui
authors.initials: H
authors.lastname: Sun
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Xu
authors.forename: Charles
authors.initials: C
authors.lastname: Rodarte
authors.forename: Alan B
authors.initials: AB
authors.lastname: Moy
authors.forename: Mack H
authors.initials: MH
authors.lastname: Wu
authors.forename: Sarah Y
authors.initials: SY
authors.lastname: Yuan
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives / Q000494:pharmacology; D000199:Actins / Q000378:metabolism; D000418:Albumins / Q000493:pharmacokinetics; D000577:Amides / Q000494:pharmacology; D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D003331:Coronary Vessels / Q000378:metabolism*; D017097:Electric Impedance; D042783:Endothelial Cells / Q000378:metabolism / Q000502:physiology; D004730:Endothelium, Vascular / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins; D007537:Isometric Contraction / Q000187:drug effects / Q000502:physiology; D008297:Male; D018994:Myosin Light Chains / Q000378:metabolism; D018375:Neutrophil Activation; D010766:Phosphorylation; D011108:Polymers / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000494:pharmacology / Q000502:physiology*; D011725:Pyridines / Q000494:pharmacology; D013552:Swine; D014661:Vasoconstriction / Q000187:drug effects / Q000502:physiology*; D014699:Venules / Q000378:metabolism*; D054460:rho-Associated Kinases
chemical_list: C459092:2-methyl-1-((4-methyl-5-isoquinolinyl)sulfonyl)homopiperazine; D000199:Actins; D000418:Albumins; D000577:Amides; D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D018994:Myosin Light Chains; D011108:Polymers; D011725:Pyridines; C108830:Y 27632; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases