Effect of cGMP on lung microvascular endothelial barrier dysfunction following hydrogen peroxide.
The authors determined the effect of cyclic guanosine 3',5'-monophosphate (cGMP) on hydrogen peroxide (H(2)O(2))-induced barrier dysfunction in bovine lung microvascular endothelial cell (BLMVEC) monolayers and compared the results to bovine pulmonary artery endothelial cells (BPAECs). In BLMVECs, H(2)O(2) (250 microM) caused a 31.9% +/- 4.8% decrease in transendothelial electrical resistance (TER) associated with increased actin stress fiber formation, intercellular gaps, and intracellular calcium concentration ([Ca(2+)](i)). The cGMP analogue 8-(p-chlorophenylthio)-cGMP (8p-CPT-cGMP; 30 or 50 microM) prevented the H(2)O(2)-induced decrease in TER (p <.001) as well as the cytoskeletal rearrangement and intercellular gap formation. 8-pCPT-cGMP (50 microM) attenuated the peak (418.8 +/- 42.1 versus 665.2 +/- 38.0 nmol/L; p <.001) and eliminated the sustained increase in [Ca(2+)](i) (193.5 +/- 21.3 versus 418.8 +/- 42.1 nmol/L; p <.001) caused by H(2)O(2). 8-pCPT-cGMP also increased TER (14.2% +/- 2.2%; p <.05) and decreased [Ca(2+)](i) (201.2 +/- 12.5 vs. 214.4 +/- 12.1 nmol/L; p <.03) before H(2)O(2). In BPAECs, 8p-CPT-cGMP significantly attenuated H(2)O(2)-induced increases in permeability and [Ca(2+)](i) but less effectively than in BLMVECs. These results suggest that in BLMVECs, cGMP countered the adverse effects of H(2)O(2) on barrier function by preventing actin cytoskeletal rearrangement and attenuating the increase in [Ca(2+)](i).
Pearse
David B
DB
Department of Medicine, Division of Pulmonary and Critical Care Medicine, Johns Hopkins Medical Institutions, Baltimore, Maryland 21224, USA. dpearse@jhmi.edu
Shimoda
Larissa A
LA
Verin
Alexander D
AD
Bogatcheva
Natalia
N
Moon
Cheil
C
Ronnett
Gabriele V
GV
Welsh
Laura E
LE
Becker
Patrice M
PM
D000199:Actins / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects / Q000502:physiology*; D002417:Cattle; D002460:Cell Line; D006152:Cyclic GMP / Q000494:pharmacology*; D017097:Electric Impedance; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D017629:Gap Junctions / Q000187:drug effects / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D008168:Lung / Q000098:blood supply* / Q000166:cytology; D016877:Oxidants / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
D000199:Actins; D016877:Oxidants; D006861:Hydrogen Peroxide; D006152:Cyclic GMP; D002118:Calcium