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CV-159, a unique dihydropyridine derivative, prevents TNF-induced inflammatory responses in human umbilical vein endothelial cells.
CV-159, a 1,4-dihydropyridine derivative, has Ca(2+) antagonistic and anti-calmodulin actions. An early feature of atherosclerosis is vascular endothelial inflammatory change. We examined whether CV-159 has protective effects against endothelial inflammatory responses. After pretreatment of human umbilical vein endothelial cells (ECs) with CV-159 (10 microM, 30 min), TNF-alpha (10 ng/ml) was applied for 20 min or 24 h. Expressions of inflammatory markers and activation of inflammatory signal molecules were examined by Western blotting. Reactive oxygen species (ROS) generation was examined by using 2',7'-dichlorodihydrofluorescein diacetate. CV-159 inhibited TNF (24 h)-induced expression of e-selectin but not vascular cell adhesion molecule-1 and intercellular adhesion molecule-1. CV-159 inhibited TNF (20 min)-induced phosphorylation of JNK, p38, and NF-kappaB p65 (Ser536). A JNK inhibitor, SP600125, and a p38 inhibitor, SB203580, inhibited TNF-induced e-selectin expression. An antioxidant drug, N-acetyl-L-cysteine (NAC), inhibited TNF-induced e-selectin expression. NAC inhibited TNF-induced phosphorylation of JNK and p38 but not NF-kappaB. CV-159 inhibited TNF-induced ROS generation. Our results indicate that in ECs CV-159 specifically inhibits TNF-induced e-selectin expression through inhibition of JNK, p38, and NF-kappaB phosphorylation. It is suggested that CV-159 prevents activation of JNK and p38 through inhibition of ROS, while it prevents activation of NF-kappaB via a ROS-independent manner.
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