Decrease of Intracellular Chloride Concentration Promotes Endothelial Cell Inflammation by Activating Nuclear Factor-&kgr;B Pathway

Recent evidence suggested that ClC-3 channel/antiporter is involved in regulation of nuclear factor (NF)-&kgr;B activation. However, the mechanism explaining how ClC-3 modulates NF-&kgr;B signaling is not well understood. We hypothesized that ClC-3-dependent alteration of intracellular chloride concentration ([Cl−]i) underlies the effect of ClC-3 on NF-&kgr;B activity in endothelial cells. Here, we found that reduction of [Cl−]i increased tumor necrosis factor-&agr; (TNF&agr;)-induced expression of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 and adhesion of monocytes to endothelial cells (P<0.05; n=6). In Cl− reduced solutions, TNF&agr;-evoked I&kgr;B kinase complex &bgr; and inhibitors of &kgr;B&agr; phosphorylation, inhibitors of &kgr;B&agr; degradation, and NF-&kgr;B nuclear translocation were enhanced. In addition, TNF&agr; and interleukin 1&bgr; could activate an outward rectifying Cl− current in human umbilical vein endothelial cells and mouse aortic endothelial cells. Knockdown or genetic deletion of ClC-3 inhibited or abolished this Cl− conductance. Moreover, Cl− channel blockers, ClC-3 knockdown or knockout remarkably reduced TNF&agr;-induced intercellular adhesion molecule 1 and vascular cell adhesion molecule 1expression, monocytes to endothelial cell adhesion, and NF-&kgr;B activation (P<0.01; n=6). Furthermore, TNF&agr;-induced vascular inflammation and neutrophil infiltration into the lung and liver were obviously attenuated in ClC-3 knockout mice (P<0.01; n=7). Our results demonstrated that decrease of [Cl−]i induced by ClC-3-dependent Cl− efflux promotes NF-&kgr;B activation and thus potentiates TNF&agr;-induced vascular inflammation, suggesting that inhibition of ClC-3-dependent Cl− current or modification of intracellular Cl− content may be a novel therapeutic approach for inflammatory diseases.

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