Rho-kinase contributes to hypoxia/reoxygenation-induced cerebral endothelial dysfunction.

Hypoxia/reoxygenation (H/R) in vitro induced cerebral endothelial dysfunction is mediated by superoxide production. However, the intracellular pathways involved remain unclear. The present study was designed to investigate the involvement of Rho-kinase and its interaction with nitric oxide (NO) in cerebral endothelial dysfunction after H/R. Arterial diameter and intraluminal pressure were simultaneously measured in vitro on rat posterior cerebral arteries. Vascular NO production was determined by measuring stable NO metabolites nitrate/nitrite. H/R selectively inhibited cerebral vasodilation to the endothelium-dependent agonist acetylcholine (ACh, 0.01 to 10 micromol/L) and caused NO deficiency. H/R-impaired vasodilation to ACh was reversed by Y27632 (1 micromol/L), a specific inhibitor of Rho-kinase, but not by chelerythrine (1 micromol/L), a selective inhibitor of protein kinase C. Y27632 had no protective effect in the presence of N-nitro-L-arginine methyl ester (L-NAME; 100 micromol/L), a specific endothelial NO synthase inhibitor. L-NAME (100 micromol/L) alone failed to modulate H/R-impaired vasodilation, so did L-arginine (3 mmol/L), a substrate for NO synthase. However, a stable NO donor diethylenetetra amine-NONOate (5 micromol/L) normalized H/R-impaired dilation to ACh. In conclusion, H/R-induced endothelial dysfunction is associated with activation of Rho-kinase-dependent pathway and NO deficiency. Pretreatment with either Y27632 or the stable NO donor profoundly prevented H/R-mediated cerebral endothelial dysfunction.