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目的:体外培养冠心病患者外周血单个核细胞来源的树突状细胞(dendritic cells,DCs),并利用基因芯片技术对冠心病患者外周血培养的DC功能基因表达谱进行研究。方法:分别配对筛选急性冠状动脉综合征患者(ACS)及冠状动脉正常的胸痛综合征患者各5例,将人外周血分离的单个核细胞加入包含rhGM-CSF(20μg/L)和rhIL-4(20μg/L)的培养基中培养,使其分化为DCs。再用基因芯片技术检测DC特异性功能蛋白基因表达的变化。结果:ACS组功能蛋白表达基因中有5个基因呈明显上升,分别是G1P2、G1P3、IFIT4、IL-1β和MX1,这些都是高度相关的干扰素诱导蛋白基因。有17个基因呈明显下降表达,分别是ACPP、AIM2、ATM、CCR1、CCR5、CD1C、FCGR3A、IFI16、IL-16、IL-18、LY75、MAP4K3、TAP1、TAP2、TLR1、TNFRSF6和VCL等,分别属于抗原识别受体、细胞趋化因子受体、细胞因子和细胞内信号传导系统。结论:冠心病患者外周血单个核细胞来源的DC不但在形态和表型上存在明显差异,而且在功能蛋白表达方面也存在显著变化。
OBJECTIVE: To culture dendritic cells (DCs) derived from peripheral blood mononuclear cells in patients with coronary heart disease in vitro and study the gene expression profiles of DCs in peripheral blood of patients with coronary heart disease using gene chip technique. METHODS: Five patients with acute coronary syndrome (ACS) and normal chest pain syndrome were enrolled in this study. Five mononuclear cells isolated from human peripheral blood were added into the culture media containing rhGM-CSF (20μg / L) and rhIL-4 (20 μg / L) to differentiate into DCs. Gene chip technology was used to detect the changes of DC-specific functional protein gene expression. Results: Five genes of functional protein expressed in ACS group were significantly increased, which were G1P2, G1P3, IFIT4, IL-1β and MX1, which were all highly correlated interferon induced protein genes. Seventeen genes were significantly down-regulated and were ACPP, AIM2, ATM, CCR1, CCR5, CD1C, FCGR3A, IFI16, IL- 16, IL- 18, LY75, MAP4K3, TAP1, TAP2, TLR1, TNFRSF6 and VCL, Respectively belong to antigen recognition receptors, chemokine receptors, cytokines and intracellular signaling systems. Conclusion: DCs derived from peripheral blood mononuclear cells of patients with coronary heart disease have not only obvious morphological and phenotypic differences but also significant changes in functional protein expression.