IL-8对OX-LDL诱导的RAW264.7株作用的研究

The role of interleukin-8 in OX-LDL-induced U264.7 cells

目的探讨白细胞介素8(IL-8)对OX-LDL诱导的小鼠单核巨噬细胞264.7细胞株(RAW264.7)的作用。方法以IL-8作用于OX-LDL诱导RAW264.7细胞株,48 h后收集细胞和上清培养液,分别用流式细胞仪检测细胞表位CD14、CD34、CD86及其早、晚期凋亡率;用RT-PCR法检测细胞内IL-8 mRNA的表达水平;用Western bot检测IL-8 mRNA表达蛋白的量;用ELISA方法检测上清液IL-1β、IL-10的量。结果与对照组相比,实验组的CD14、CD34表位表达及晚期凋亡明显增多(P<0.05),而对CD86、早期凋亡影响不明显(P>0.05),IL-8 mRNA及其表达的蛋白量明显升高,上清培养液中IL-1β增多(P<0.05),IL-10减少(P<0.05)。结论 IL-8致动脉粥样硬化(AS)作用可能是部分通过上调单核巨噬细胞表面CD14、CD34表位表达,促进IL-8 mRNA及其表达蛋白的量,促进IL-1β、抑制IL-10的分泌,并促进其晚期凋亡,参与AS的炎症反应。

Objective To explore the role of IL-8 in OX-LDL-induced U264.7 cells.Methods OX-LDL-induced RAW264.7 cells were stimulated by IL-8 for 48 hours,Cells and supernatants were collected.CD14,CD34,CD86 and the early/ terminal apoptotic rate of U264.7cells were examined by flow cytometry,and the expression of IL-8 mRNA and protein were determined by real-time quantitative RT-PCR and Western blot,respectively.The levels of IL-1β,IL-10 in the supernatants were determined by ELISA.Results Compared with the control group,the levels of CD14,CD34 and CD86 in experimental group were significantly increased（P0.05）.The rate of terminal apoptosis was significantly increased（P0.05）,but the rate of early apoptosis and CD86 level were not affected（P0.05）.The levels of IL-8 mRNA and protein were significantly increased.The levels of IL-1β increased and IL-10 decreased significantly in the supernatants compared with those in the control group（P0.05）.Conclusion The proatherogenic effect of IL-8 in atherosclerosis（AS） may be partly through up-regulating CD14,CD34 expression in macrophages by increasing IL-8 mRNA and protein,promoting its secretion of IL-1β and suppressing IL-10 level,boosting macrophage terminal apoptosis to participate in the inflammatory responses of AS.