藏红花素对白血病患儿骨髓树突状细胞体外增殖作用的影响

Effect of Crocin on Culture and Proliferation of Dendritic Cells Derived from Children Acute Leukemia Blood marrow In Vitro

目的:探讨藏红花素(crocin)对白血病患儿骨髓树突状细胞体外增殖作用的影响。方法:采用Ficoll-Hypaque法分离急性白血病患者骨髓液单个核细胞,将本实验分为六组。培养至第9天,在倒置显微镜下观察各组细胞形态,对各组细胞进行计数,并行瑞氏-姬姆萨染液染色,在油镜下观察典型树突状细胞的形态,应用流式细胞仪检测各组细胞免疫表型。结果:结果表明,实验各组均得到一定数量典型的DC,对照组中未见,实验各组细胞数目明显高于对照组,差异有显著统计学意义(P<0.01)。在加入细胞因子的各组中,以E组即加入藏红花素1.25mg/ml组细胞数最高,而C组细胞数与D组细胞数比较无显著差异(P>0.05)。培养至第9天,流式细胞仪免疫分析显示C、D、E、F各组CD1a+、CD83+、HLA-DR+细胞比例明显高于A、B两组(P<0.01),差异有显著统计学意义,A、B两组之间比较差异无统计学意义(P>0.05)。D、F组与C组比较,差异无统计学意义(P>0.05)。E组与C、D、F组比较,差异有统计学意义(P<0.05)。结论:藏红花素能促进树突状细胞增殖,其促进树突状细胞增殖的能力明显弱于细胞因子的联合作用。藏红花素协同细胞因子促进树突状细胞的成熟具有浓度依赖性,在藏红花素浓度为1.25mg/ml时作用最强,而浓度过高或过低时均未表现出明显的促进作用。

Objective： To investigate the effect of crocin on the culture and proliferation and immunifaction of dendritic cells（DC） derived from leukemia children acute leukemia blood marrow in vitro.Methods： The mononuclear cells were isolated from blood marrow of Leukemia children by the method of Ficoll-Hypaque.The experiment were divided into six groups.The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture,and some harvest cells were stained with Wright-Giemsa,then observed and photographed under the oil immersion objective.Results： The test groups all attained a certain amount of typical DC except the control group.The numbers of test groups were higher than the control group（P0.01）.The numbers between C group and D group were no significant difference（P0.05）.Cultured for 9 days,the rates of CD1a＋,CD83＋ and HLA-DR＋ in group C,D,E and F were higher than that of group A and B（P0.01）.There was no significant difference between A and B group（P0.05）.There was no significant difference between group D,F and C（P0.05）.There was significant difference between E and C,D,F group（P0.05）.Conclusion： Crocin can promote the growth and transformation of dendritic cells.The power of the crocin is lower than that of the isola-tion of the rhGM-CSF,rhIL-4 and rhTNF-α.Crocin synergistic cytokines promote the maturation of dendritic cells in a concentration-de-pendent,the effect is strongest when the concentration is 1.25 mg/ml.When the concentration is too high or too low,there is no obvious role in promoting.