华蟾素对正常人外周血来源树突状细胞影响.docVIP

华蟾素对正常人外周血来源树突状细胞影响 　【摘要】 目的：从树突状细胞(DCs)的分子细胞水平初步探讨华蟾素提高机体免疫力的作用机制。方法：分离正常人外周血单核细胞，以GM-CSF+IL-4培养诱导DCs，采用自身配对设计分成对照组和华蟾素组（DCs培养48小时后加入华蟾素）。培养第7天镜下观察两组细胞形态的异同，用流式细胞仪检测DCs表面分子的表达水平，用CCK8检测DCs刺激同种异体淋巴细胞增殖的能力，用ELISA法检测DCs培养液上清中IL-12的水平。结果：华蟾素组DCs与对照组比较细胞形态相对成熟，表面分子表达水平增高，刺激同种异体淋巴细胞增殖能力和IL-12分泌能力增强。结论：华蟾素能增强正常人DCs的功能。 【关键词】 树突状细胞 华蟾素 外周血 流式细胞仪 酶联免疫吸附试验 [Abstract] Objective:To study the effect of Cinobufotalin on the morphous，phenotype and function of dendritic cells derived from peripheral blood of health adults, and to explain the mechanism of action of Cinobufotalin from the molecule and cell levels of DCs. Methods:Mononuclear cells of healthy adults were isolated from peripheral blood, and were cultured to induce DCs with the combination of GM-CSF and IL-4. Using self matched-pairs design two groups were divided in the experiment: controls and Cinobufotalin（Cinobufotalin was added to the culture media after the cells had been incubated for 48 hours）. On the 7th day, DCs of 2 groups were observed to compare the differences of the morphous, the expression of surface molecules on DCs were detected by flow cytometer; the DCs? capacities of stimulating allogeneic lymphocyte proliferation was detected by cell count kit 8(CCK8), and the concentration of IL-12 in the supernatants of DCs? cultures was detected by ELISA. Results: compared with the controls, the DCs incubated with Cinobufotalin had more mature state, had higher expression of surface molecules, and higher capacities of stimulating allogeneic lymphocyte proliferation and secreting IL-12. Conclusion:Cinobufotalin can enhance the function of DCs of healthy adults. [Key words] Dendritic cell; Cinobufotalin； Peripheral blood； Flow cytometer； Enzyme-linked immunosorbent assay 华蟾素(cinobufotalin)是由中华大蟾蜍阴干的全皮为原料提取的水溶性制剂，应用于临床治疗乙肝及肝癌已有20多年的历史。临床应用表明其能显著提高机体的免疫功能[1，2]。但有关华蟾素作用机制的研究很少，本课题通过研究华蟾素对正常人外周血来源的树突状细胞(DCs)形态、表型和功能的影响，从DCs的分子细胞水平初步探讨华蟾素提高机体免疫功能的作用机制。 1 材料与方法 1.1 材料 1.1.1 细胞因子和单克隆抗体 rhGM-CSF(北京宝赛生物技术有限公司)、rhIL-