P38α subtype is a potential target to inhibit eNOS activity and NO production in human endothelial cells

Abstract

Human endothelial nitric oxide synthase (eNOS) activity is important for maintaining blood pressure homeostasis and vascular integrity through nitric oxide (NO).The in vitro study aimed at investigating a role of p38α signaling in modulating NO production in human umbilical vein endothelial cell-12 (HUVEC-12). Consistent with the stimulation of lipopolysaccharide (LPS) or tumor necrosis factor (TNF)-α, the over-expression of p38α markedly down-regulated the eNOS promoter activity in HUVEC-12, which could be reversed by its negative mutant p38α (AF) or p38-specific inhibitor SB203580. Compared to the stimulation of LPS or TNF-α, p38α-targeting siRNA decreased the expressions of phosphorylated and non-phosphorylated p38α, and increased the promoter activity, an eNOS mRNA level and a phosphorylated eNOS protein expression with the enhancement of NO, which could be abrogated by the scrambled siRNA. The in situ eNOS protein expression in the cells treated by p38α-targeting siRNA was also higher than that of the control, following the corresponding attenuation of a p38 level, and mainly localized in the inner membrane and cytoplasm. These results indicate that the p38α subtype may be a potential target to down-regulate the eNOS activity and NO production in human endothelial cells.