缬沙坦可阻断酒精诱导的、Toll样受体2信号介导的人血管内皮细胞炎症反应。

Valsartan blocked alcohol-induced, Toll-like receptor 2 signaling-mediated inflammation in human vascular endothelial cells.

作者信息

Wang Yushu, Li Yi, Shen Qingyu, Li Xiangpen, Lu Juan, Li Xiangping, Yin Deling, Peng Ying

机构信息

Department of Neurology, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, China.

出版信息

Alcohol Clin Exp Res. 2014 Oct;38(10):2529-40. doi: 10.1111/acer.12532.

DOI:10.1111/acer.12532

PMID:25346502

Abstract

BACKGROUND

Alcohol consumption induces inflammatory damage in vessels, and the underlying mechanism is unclear. Valsartan, as one of the angiotensin receptor blockers (ARBs), plays a role in the inhibition of inflammatory reactions in vascular dysfunction. This study is to investigate the role of Toll-like receptor 2 (TLR2) in alcohol-induced inflammatory damage in vascular endothelial cells in vitro and to explore the protective effect of valsartan on alcohol-induced and TLR2-mediated inflammatory damage.

METHODS

The human umbilical vein cell line (EA.hy926) were exposed to alcohol at 0 to 80 mM for 0 to 48 hours with or without valsartan pretreatment. The expression of TLR2 signaling, including TLR2, tumor necrosis factor receptor associated factor 6 (TRAF-6) and nuclear factor kappa B (NF-κB) p65 were detected by Western blot. The levels of proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), were determined by ELISA. To confirm the role of TLR2, we functionally up-regulated or down-regulated TLR2 by using TLR2 agonist or TLR2 small interfering RNA (siRNA), respectively. To further investigate the mechanism of alcohol on renin-angiotensin system, we detected the expression of angiotensin II receptor type 1 (AGTR1) in protein levels.

RESULTS

The expression of TLR2, TRAF-6, NF-κB p65, and the proinflammatory cytokines, TNF-α and IL-6, were significantly increased after alcohol exposure in EA.hy926 endothelial cells. This was enhanced by TLR2 agonist, and was inhibited by TLR2 siRNA transfection. The pretreatment of valsartan resulted in an inhibition of TLR2 signaling and proinflammatory cytokines. The expression of AGTR1 was up-regulated after alcohol exposure, and was blocked by valsartan pretreatment.

CONCLUSIONS

TLR2 signaling-mediated alcohol induced inflammatory response in human vascular epithelial cells in vitro, which was inhibited by valsartan.

摘要

背景

酒精摄入会引发血管的炎症性损伤，但其潜在机制尚不清楚。缬沙坦作为血管紧张素受体阻滞剂（ARB）之一，在抑制血管功能障碍中的炎症反应方面发挥作用。本研究旨在探讨Toll样受体2（TLR2）在酒精诱导的体外血管内皮细胞炎症损伤中的作用，并探索缬沙坦对酒精诱导的和TLR2介导的炎症损伤的保护作用。

方法

人脐静脉细胞系（EA.hy926）在有或无缬沙坦预处理的情况下，暴露于0至80 mM的酒精中0至48小时。通过蛋白质印迹法检测TLR2信号通路的表达，包括TLR2、肿瘤坏死因子受体相关因子6（TRAF - 6）和核因子κB（NF - κB）p65。通过酶联免疫吸附测定法（ELISA）测定促炎细胞因子肿瘤坏死因子-α（TNF - α）和白细胞介素-6（IL - 6）的水平。为了证实TLR2的作用，我们分别使用TLR2激动剂或TLR2小干扰RNA（siRNA）在功能上上调或下调TLR2。为了进一步研究酒精对肾素-血管紧张素系统的作用机制我们检测了血管紧张素II 1型受体（AGTR1）的蛋白水平表达。

结果

酒精暴露后，EA.hy926内皮细胞中TLR2、TRAF - 6、NF - κB p65以及促炎细胞因子TNF - α和IL - 6的表达显著增加。TLR2激动剂增强了这种增加，而TLR2 siRNA转染则抑制了这种增加。缬沙坦预处理导致TLR2信号通路和促炎细胞因子受到抑制。酒精暴露后AGTR1的表达上调，而缬沙坦预处理则阻断了这种上调。