姜黄素诱导的免疫耐受性人树突状细胞的研究

摘要

Objective:To observe the induction of human to lerogenic dendritic cells by curcum in.Methods:Peripheral blood mononu-clear cells were isolated from human peripheral blood by Fico11-Hypaque density gradient centrifugation,and then cultured in the medium in-cluding rhGM-CSF and rhIL-4 for 6 days.There are four groups:①immature DC group(imDC group),②curcum in group(Curgroup),③cur-cum in and lipopolysaccharide group(Cur+LPS group),④lipopolysaccharide group(LPS group).The DCs were analyzed by flow cytometry for the expression of CD80,CD83,CD86 and HLA-DR.Endocytosis of DCs to dextran was also amalyzed by flow cytometry.The production of IL-12 in DC culture supernatant was determined by ELISA.The proliferation of allogenetic T cells was detected by MTT method.Result:Curcum in suppressed the expression of costimulatory molecules(CD80,CD83,CD86)and the antigen presenting molecule(HLA-DR) in a does-dependent manner.There was no marked difference in the surface molecules expression of Cur+LPS group as compared with immature DCs.The positive rate of dextran uptake of Cur+LPA group was remarkably higher than in LPS group(p＜0.05).While DCs in Cur+LPS group showed the de-creased capacity to stimulate T cell proliferation as compared with that in LPS group(p＜0.05).Also the secretory amounts of IL-12 of DCs in Cur+LPS group were obviously lower than in LPS group(p＜0.05).Conclusion:Curcum in can inhibit the maturation of human dendritic cells,and lead to the tolerogenic dendritic cells.%目的:探讨姜黄素(Curcumin)诱导免疫耐受性人树突状细胞(Dendritic cell,DC)的效果.方法:聚蔗糖-泛影葡胺(ficoll-hypaque,F-H)密度梯度离心法获得健康人外周血单个核细胞(PBMC),在含有重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)、重组人白介素4(rhIL-4)的培养基中培养6天.实验共分四组:①未成熟DC组(imDC组)、②姜黄素组(Curcumin,Cur组)、③姜黄素+脂多糖组(Cur+LPS组)、④脂多糖组(LPS组).流式细胞术检测DC表型CD80、CD86、CD83及HLA-DR的表达情况和DC吞噬葡聚糖的能力,酶联免疫吸附法(ELISA)检测DC分泌白介素-12(IL-12)的能力,混合淋巴细胞反应(MLR)检测DC刺激同种异体T淋巴细胞增殖的能力.结果:Cur能够显著抑制DC共刺激分子CD80、CD86、CD83及HLA-DR的表达,并呈剂量依赖性,与imDC组比较差异无统计学意义;与LPS组比较,Cur+LPS组DC吞噬葡聚糖的阳性细胞百分率显著提高(P＜0 05),而刺激同种异体T淋巴细胞增殖的能力则显著下降(P＜0.05),IL-12的分泌量也显著下降(P＜0.05).结论:姜黄素能够抑制人树突状细胞的成熟,从而获得免疫耐受性的人树突状细胞.