氯化锦葵色素-3-半乳糖苷对TNF-α诱导内皮细胞炎症的抑制作用

Inhibitory Effect of Malvidin-3-Galactoside Chloride on TNF-α-Induced Inflammation in Endothelial Cells

目的:研究氯化锦葵色素-3-半乳糖苷对细胞肿瘤坏死因子(tumour necrosis factor-α,TNF-α)诱导人脐静脉内皮细胞(human vascular umbilical endothelial cells,HUVEC)所产生炎症的抑制作用。方法:设立分组,空白组(二甲基亚砜)、10μg/L TNF-α刺激组、浓度为1、10、50、100μmol/L氯化锦葵色素-3-半乳糖苷+10μg/L TNF-α刺激组,通过酶联免疫吸附剂测定方法检测细胞上清中可溶性细胞间黏附因子(intercellular cell adhesion molecule-1,ICAM-1)蛋白含量的变化;用实时荧光定量聚合酶链式反应检测细胞中ICAM-1 mRNA表达量变化;Western blotting检测ICAM-1及核因子κB抑制蛋白(IκBα)蛋白含量的变化;通过免疫荧光方法测定核转录因子κB(nuclear transcription factor-κB,NF-κB)核异位的变化。结果:氯化锦葵色素-3-半乳糖苷呈浓度依赖性的抑制TNF-α诱导HUVEC相应蛋白表达,同时结果显示抑制作用与NF-κB信号通路有关。结论:氯化锦葵色素-3-半乳糖苷抑制TNF-α诱导HUVEC的炎症反应。一般心血管疾病产生的原因中包括因促炎症因子引起的内皮功能障碍及产生的炎症,氯化锦葵色素-3-半乳糖苷的抗炎症作用为防治心血管疾病提供了一定的理论基础。

Objective： The effect of malvidin-3-galactoside chloride （Mv-3-gal-C1） on TNF-α-induced inflammation in human umbilical vein endothelial cells （HUVEC） was investigated in this study. Methods： HUVEC were treated with TNF-α （10 μg/L） alone and combined with Mv-3-gal-C1 （1, 10, 50 and 100 μmol/L）, respectively. DMSO was used as control. The soluble ICAM-1 protein content in cell supernatant was detected by ELISA. The change in ICAM-1 mRNA expression level in ceils was detected by RT-PCR. The expression of ICAM-1 and IKβα was assessed by Western blotting. The activity of NF-KB was evaluated by immunofluorescence. Results： Mv-3-gal-C1 appeared to specifically down-regulate the TNF-a-induced cell surface expression of ICAM-1 and ICAM-1 mRNA expression and protein release from HUVEC cells in a concentration-dependent manner. The inhibitory effect of Mv-3-gal-C1 on protein expression might be associated with the suppression of NF-kB in HUVECs. Conclusion： Mv-3-gal-C1 can inhibit TNF-α-induced inflammation in HUVECs. The anti-inflammatory effect of Mv-3-gal-C1 provides a theoretical basis for preventing and treating pro-inflammatory cytokines- induced endothelial dysfunction and cardiovascular diseases resulting from inflammation.