Carbachol inhibits TNF-α-induced endothelial barrier dysfunction through alpha 7 nicotinic receptors

摘要

Aim:To test whether carbachol can influence endothelial barrier dysfunction induced by tumor necrosis factor(TNF)-αand whether the alpha 7 nicotinic receptor can mediate this process.Methods:Rat cardiac microvascular endothelial cells were exposed to carbachol followed by TNF-α treatment in the presence or the absence of α-bungarotoxin(an antagonist of the alpha 7 nicotinic receptor).Permeability of endothelial cells cultured on Transwell filters was assayed using FITC-albumin.F-actin was stained with FITC-phalloidin.Expression of vascular endothelial cadherin,intercellular adhesion molecule 1(ICAM-1),phosphor-ERK1/2 and phosphor-JNK was detected using Western blot.Results:Carbachol(2μmol/L-2 mmol/L)prevented increase in endothelial cell permeability induced by TNF-α(500 ng/mL)in a dose-dependent manner.Further,it attenuated the down-regulation of vascular endothelial cadherin and the up-regulation of ICAM-1 induced by TNF-α.In addition,treatment of endothelial cells with carbachol decreased phosphor-ERK1/2 and phosphor-JNK.These effects of carbachol were blocked by α-bungarotoxin 3 μg/mL.Conclusion:These data suggest that the inhibitory effect of carbachol on TNF-α-induced endothelial barrier dysfunction mediated by the alpha 7 nicotinic receptor.