Barrier dysfunction and RhoA activation are blunted by homocysteine and adenosine in pulmonary endothelium.
RhoA GTPases modulate endothelial permeability. We have previously shown that adenosine and homocysteine enhance basal barrier function in pulmonary artery endothelial cells by a mechanism involving diminution of RhoA carboxyl methylation and activity. In the current study, we investigated the effects of adenosine and homocysteine on endothelial monolayer permeability in cultured monolayers. Adenosine and homocysteine significantly attenuated thrombin-induced endothelial barrier dysfunction and intercellular gap formation. We found significantly diminished RhoA associated with the membrane subcellular fraction in endothelial cells pretreated with adenosine and homocysteine, compared with vehicle-treated endothelial cells. Additionally, adenosine and homocysteine significantly blunted RhoA activation following thrombin exposure. Incubation with adenosine and homocysteine also enhanced in vitro interactions between RhoA and RhoGDI, as well as subcellular translocation of p190RhoGAP to the cytosol. These data demonstrate that elevated intracellular concentrations of homocysteine and adenosine enhance endothelial barrier function in cultured endothelial cells isolated from the main pulmonary artery and lung microvasculature, suggesting a potentially protective effect against pulmonary edema in response to lung injury. We speculate that homocysteine and adenosine modulate the level of endothelial barrier dysfunction through modulation of RhoA posttranslational processing resulting in diminished GTPase activity through altered interactions with modulators of RhoA activation.
Harrington
Elizabeth O
EO
Pulmonary Vascular Research Laboratory, Providence Veterans Affairs Medical Center, Providence, RI 02908, USA.
Newton
Julie
J
Morin
Nicole
N
Rounds
Sharon
S
D000241:Adenosine / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D021961:Cell Membrane Structures / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D006710:Homocysteine / Q000494:pharmacology*; D007700:Kinetics; D011652:Pulmonary Circulation / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013917:Thrombin / Q000502:physiology; D020742:rhoA GTP-Binding Protein / Q000187:drug effects / Q000502:physiology*
D006710:Homocysteine; D013917:Thrombin; D020742:rhoA GTP-Binding Protein; D000241:Adenosine