白藜芦醇对人外周血单核-巨噬细胞CCR5表达的影响.DOCVIP

临床医学论文-白藜芦醇对人外周血单核/巨噬细胞CCR5表达的影响 【摘要】? 目的： 观察白藜芦醇对人外周血单核/巨噬细胞CC型趋化因子受体5(CCR5) 表达的调节作用。方法： 采用Ficoll-Hypaque密度梯度离心法分离人外周血单个核细胞，再经贴壁法纯化单核/巨噬细胞。采用IFN-γ（1×105U/L)诱导单核/巨噬细胞表达CCR5，再分别加入不同浓度的白藜芦醇(0.5，5，25，50和100 μmol/L)进行干预。培养24 h后收集细胞，RT-PCR法检测外周血单核/巨噬细胞CCR5 mRNA表达水平；流式检测单核/巨噬细胞CCR5表达阳性率。同时将CCR5荧光素酶报告基因（pGL3-Basic-CCR5）转染各组细胞，检测各转染组的荧光素酶相对活性。结果： 中、高浓度白藜芦醇处理组（25,50和100 μmol/L）的CCR5 mRNA表达水平、CCR5阳性细胞率和CCR5-luc报告基因的荧光素酶相对活性比对照组均有所降低，低浓度白藜芦醇处理（0.5和5 μmol/L）对单核/巨噬细胞CCR5的表达无明显影响。结论： 中、高浓度白藜芦醇可抑制外周血单核/巨噬细胞CCR5的表达。 【关键词】? 白藜芦醇； 单核/巨噬细胞； 巨噬细胞移动抑制因子； 报告基因； 趋化因子受体 　　［Abstract］? Objective: To study regulating efforts of resveratrol on the expression of CC-chemokine receptor-5(CCR5)? in human peripheral monocyte/macrophage.Methods： Mononuclear cells were obtained from human peripheral blood by the method of Ficoll-Hypaque density gradient centrifugation. Macrophages were enriched by adherence. IFN-γ(1×105 U/L) were added into the medium to induce the cells expressing CCR5. Different concentration of RES(0.5， 5, 25, 50 and 100? μmol/L) were applied to the cells at the same time. Monocyte/macrophage were collected after cultured for 24 h, the expression level of CCR5 mRNA was detcted by RT-PCR and CCR5 positive cell rate was assayed by flow cytometry. CCR5 report gene were transfected into each group that mentioned above, the relative luciferase activity of CR5 report gene was tested by kit.? Results： The expression of CCR5, rates of CCR5 positive cells and relative activity of CCR5 report gene was significantly decreased in RES（25,50 and 100? μmol/L）treated group compared with control group, while there were no significant difference between control group and RES 0.5 and 5? μmol/L treated group.? Conclusion： Midst and high concentration of resveratrol have the efforts to inhibit? the expression? of? CCR5? in human peripheral monocyte/macrophage. 　　［Key words］? resveratrol；? monocyte；? macrophage；? CCR5；? report gene;? chemokine receptor 　　白藜芦醇(Resveratrol, RES, 3,5,4′-三羟基-均二苯代乙烯)是一种主要由