目的 探讨炎症因子白介素-1β(IL-1β)是否通过α1,3-岩藻糖基转移酶Ⅶ(FucT-Ⅶ)对血管内皮细胞中功能蛋白糖基化修饰调控单核细胞与内皮细胞的黏附作用,为揭示炎症因子损伤血管内皮细胞的分子机制奠定基础.方法 建立不同浓度IL-1β损伤EA.hy926血管内皮细胞的模型,通过与荧光标记的THP-1单核细胞共培养,观察细胞间的黏附作用;采用Real time-PCR及Western blot检测EA.hy926细胞中FucT-Ⅶ差异表达;用蛋白免疫印迹法检测细胞内sLex糖支链蛋白差异表达;运用siRNA干扰FucT-Ⅶ基因,观察细胞间黏附的变化.结果 IL-1β可以促进内皮细胞与单核细胞的黏附,同时内皮细胞中FucT-Ⅶ表达上调;sLex糖支链蛋白表达也上调.当干扰内皮细胞中FucT-Ⅶ基因后,细胞间的黏附能力明显下降,且sLex糖支链蛋白表达下调.结论 IL-1β可能通过调控EA.hy926细胞中FucT-Ⅶ的糖基化修饰作用增加其对单核细胞的黏附能力,这可能是炎症早期发生时影响血管内皮功能新的重要分子机制之一.%Aim To investigate whether interleukin 1βthelial cells and monocytes by Alpha 1,3-fucosyltrans-ferase Ⅶ ( FucT-Ⅶ ) modificating functional proteins glycosylation in vascular endothelial cells. Methods This study took EA. hy926 vascular endothelial cells as the object. An injury model was created by different concentrations of IL-1β treatment in EA. hy926 cells. The adhesion between cells was observed by fluorescence-labeled THP-1 monocytes co-cultured with vascular endothelial cells. Then the levels of FucT-Ⅶ mRNA and sialyl-Lewis X( sLex ) protein were detected by real-time PCR and Western blot. FucT-Ⅶ protein was down-regulated by siRNA, and then the cells adhesion was measured by 3D-culture. Results IL-1βregulates the adhesion between human vascular endo-treatment obviously enhanced EA. hy926 adhesion to THP-1. Meanwhile, the levels of FucT-YB and sLex were increased apparently. Interference of FucT-VH mRNA in EA. hy926 cell reduced fucosylation of membrane proteins and at last inhibited cell adhesion. Conclusions IL-1β maybe through up-regulating FucT-Ⅶ induced functional proteins glycosylation, thereby enhances the adhesion of monocytes to endothelial cells. This may be one of the important molecular mechanisms affecting vascular endothelial functions, which occurs in early inflammatory.
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