Effect of cGMP on lung microvascular endothelial barrier dysfunction following hydrogen peroxide.

Abstract

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).