TY - JOUR
T1 - Oxidative species increase arginase activity in endothelial cells through the RhoA/Rho kinase pathway
AU - Chandra, S.
AU - Romero, M. J.
AU - Shatanawi, A.
AU - Alkilany, A. M.
AU - Caldwell, R. B.
AU - Caldwell, R. William
PY - 2012/1
Y1 - 2012/1
N2 - Background and Purpose NO produced by endothelial NOS is needed for normal vascular function. During diabetes, aging and hypertension, elevated levels of arginase can compete with NOS for available l-arginine, reducing NO and increasing superoxide (O 2 .-) production via NOS uncoupling. Elevated O 2 .- combines with NO to form peroxynitrite (ONOO -), further reducing NO. Oxidative species increase arginase activity, but the mechanism(s) involved are not known. Our study determined the mechanism involved in peroxynitrite and hydrogen peroxide-induced enhancement in endothelial arginase activity. We hypothesized that oxidative species increase arginase activity through PKC-activated RhoA/Rho kinase (ROCK) pathway. Experimental approach Arginase activity/expression was analysed in bovine aortic endothelial cells (BAEC) treated with an ONOO - generator (SIN-1) or H 2O 2. Pretreatment with inhibitors of Rho kinase (Y-27632) or PKC (Gö6976) was used to investigate the mechanism involved in arginase activation. Key Results Exposure to SIN-1 (25 μM, 24 h) or H 2O 2 (25 μM, 8 h) increased arginase I expression and arginase activity (35% and 50%, respectively), which was prevented by ROCK inhibitor, Y-27632, PKC inhibitor, Gö6976 or siRNA to p115-Rho GEF. There was an early activation of p115-Rho GEF (SIN-1, 2 h; H 2O 2, 1 h) and Rho A (SIN-1, 4 h; H 2O 2, 1 h) that was prevented by using the PKC inhibitor. Exposure to SIN-1 and H 2O 2also reduced NOS activity, which was blocked by pretreatment with p115-RhoGEF siRNA. Conclusions and Implications Our data indicate that the oxidative species ONOO - and H 2O 2 increase arginase activity/expression through PKC-mediated activation of RhoA/Rho kinase pathway.
AB - Background and Purpose NO produced by endothelial NOS is needed for normal vascular function. During diabetes, aging and hypertension, elevated levels of arginase can compete with NOS for available l-arginine, reducing NO and increasing superoxide (O 2 .-) production via NOS uncoupling. Elevated O 2 .- combines with NO to form peroxynitrite (ONOO -), further reducing NO. Oxidative species increase arginase activity, but the mechanism(s) involved are not known. Our study determined the mechanism involved in peroxynitrite and hydrogen peroxide-induced enhancement in endothelial arginase activity. We hypothesized that oxidative species increase arginase activity through PKC-activated RhoA/Rho kinase (ROCK) pathway. Experimental approach Arginase activity/expression was analysed in bovine aortic endothelial cells (BAEC) treated with an ONOO - generator (SIN-1) or H 2O 2. Pretreatment with inhibitors of Rho kinase (Y-27632) or PKC (Gö6976) was used to investigate the mechanism involved in arginase activation. Key Results Exposure to SIN-1 (25 μM, 24 h) or H 2O 2 (25 μM, 8 h) increased arginase I expression and arginase activity (35% and 50%, respectively), which was prevented by ROCK inhibitor, Y-27632, PKC inhibitor, Gö6976 or siRNA to p115-Rho GEF. There was an early activation of p115-Rho GEF (SIN-1, 2 h; H 2O 2, 1 h) and Rho A (SIN-1, 4 h; H 2O 2, 1 h) that was prevented by using the PKC inhibitor. Exposure to SIN-1 and H 2O 2also reduced NOS activity, which was blocked by pretreatment with p115-RhoGEF siRNA. Conclusions and Implications Our data indicate that the oxidative species ONOO - and H 2O 2 increase arginase activity/expression through PKC-mediated activation of RhoA/Rho kinase pathway.
KW - RhoA
KW - arginase
KW - hydrogen peroxide
KW - peroxynitrite
KW - protein kinase C
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U2 - 10.1111/j.1476-5381.2011.01584.x
DO - 10.1111/j.1476-5381.2011.01584.x
M3 - Article
C2 - 21740411
AN - SCOPUS:83755224345
SN - 0007-1188
VL - 165
SP - 506
EP - 519
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
IS - 2
ER -