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目的探讨蒙花苷对血管内皮细胞的保护作用和机制。方法 MTT法检测肿瘤坏死因子(TNF-?)和蒙花苷对人脐静脉内皮细胞(EA.hy926)活性的影响;采用TNF-?(50 ng·m L-1)刺激EA.hy926,加入不同浓度(5,15和25?mol·L-1)蒙花苷处理24 h,DAPI染色法测定EA.hy926细胞与单核细胞(THP-1)的黏附能力;Western blot法检测细胞中Toll样受体(TLR4)、核因子-?B抑制蛋白(IκBα)、核因子-?B(NF-?B)、细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)蛋白的表达;RT-PCR法检测ICAM-1、VCAM-1 m RNA的表达;ELISA法检测细胞上清液中白介素-1(IL-1)、白介素-6(IL-6)和白介素-8(IL-8)的含量。结果蒙花苷可显著抑制TNF-?刺激EA.hy926细胞与THP-1细胞间黏附作用,降低EA.hy926细胞分泌的炎症因子IL-1、IL-6和IL-8水平。Western blot和RT-PCR结果表明蒙花苷能减少EA.hy926细胞中TLR4、NF-?B、ICAM-1、VCAM-1的蛋白表达量及ICAM-1、VCAM-1 m RNA的表达量,同时能升高细胞中IκBα的蛋白表达量。结论蒙花苷能缓解血管内皮细胞炎症损伤,其作用机制可能主要是通过抑制TLR4/IκBα/NF-κB信号通路发挥效用。
Objective To investigate the protective effect and mechanism of mandarin glycoside on vascular endothelial cells. Methods MTT assay was used to detect the effects of tumor necrosis factor (TNF-α) and mandarin on human umbilical vein endothelial cells (EA.hy926). EA.hy926 was stimulated with TNF-α (50 ng · m L -1) Adiponectin at different concentrations (5, 15 and 25 mol·L -1) was treated for 24 h, and the adhesion ability of EA.hy926 cells to monocytes (THP-1) was determined by DAPI staining. The levels of Toll TLR4, NFκBα, NF-κB, ICAM-1 and VCAM The expression of ICAM-1 and VCAM-1 mRNA was detected by RT-PCR and the level of interleukin-1 (IL-1), interleukin-6 Interleukin-8 (IL-8) content. Results Monatin could significantly inhibit the adhesion between EA.hy926 cells and THP-1 cells and decrease the levels of inflammatory cytokines IL-1, IL-6 and IL-8 secreted by EA.hy926 cells. The results of Western blot and RT-PCR showed that monatin inhibited the protein expression of TLR4, NF-? B, ICAM-1 and VCAM-1 and the expression of ICAM-1 and VCAM-1 mRNA in EA.hy926 cells, At the same time can increase the IκBα protein expression in cells. Conclusions Monacolin can relieve the inflammatory injury of vascular endothelial cells. Its mechanism may be mainly through the inhibition of TLR4 / IκBα / NF-κB signaling pathway.