大黄素对体外诱导人血来源树突状细胞的影响

Effects of emodin on dendritic cells from human peripheral blood induced culture in vitro

目的探讨大黄素对体外诱导培养的人血来源树突细胞(DC)的影响。方法分离健康人外周血单核细胞,经培养后获得未成熟DC(iDC),重组人粒-巨噬细胞集落刺激因子(rhGM-CSF,106IU.L-1)和重组人白细胞介素4(rhIL-4,8×105IU.L-1)诱导获得成熟DC(mDC)。实验分为iDC组、mDC组和大黄素组,mDC组于d 5加入脂多糖(LPS,1 mg.L-1)刺激,大黄素组采用mDC在d 5经LPS刺激后于d 7加入大黄素(100 mg.L-1)共培养2 d。用倒置显微镜和电镜观察DC细胞形态,用流式细胞仪检测DC表面共刺激分子的表达水平。结果经rhGM-CSF、rhIL-4诱导和LPS刺激,培养9 d后可获得表面有丰富分叉状胞浆突起的毛刺状mDC。大黄素组DC表面突起短而少,呈iDC形态。大黄素组CD80、CD83和CD86表达率分别为(13.4±6.6)%、(9.3±2.2)%和(84.2±6.3)%,低于mDC组[分别为(39.3±8.6)%、(30.7±5.6)%和(95.4±3.2)%,P<0.01];大黄素组CD14表达率为(8.4±2.8)%显著高于mDC组的(3.7±2.3)%(P<0.01)。大黄素组HLA-DR及CD11c表达与mDC组比较无显著差异(P>0.05)。结论大黄素能干扰DC表面突起的形成和表面共刺激分子的表达。

AIM To investigate the effects of emodin on dendritic cells （DC） from human peripheral blood induced culture in vitro. METHODS Cells were separated from human peripheral blood mononuclear cells （PBMCs） to obtain immature DE （iDC） after cell training. Mature DC （mDC） were induced by recombinant human interleukin-4 （rhlL-4） and recombinant human granulocyte-macrophage colony stimulating factor （rhGM- CSF）. Three groups were involved as iDC group, mDC group and emodin group. The mDC group was stimulated by lipopolysaccharide （LPS, 1 mg＇L-1） on the fifth day, and emodin （100 mg.L^-1） was added on the seventh day in the emodin group for two days. The suspending cells were examined with inverted microscope and scanning electronic microscope. The phenotype of DCs （HLA-DR, CD80, CDa6, CD83, CD14, CD11c） were analyzed by flow cytometry. RESULTS PBMCs exhibited typical morphological characteristics of DC occurred when induced by rhGM-CSF with rhIL-4 and treated by LPS. The expression rates of CD80, CD83 and CD86 in the emodin group were （13.4±6.6） %, （9.3 ± 2.2） % and （84.2 ± 6.3） % respectively, which were significantly lower than those in the control group （（39.3 ± 8.6） %, （30.7 ± 5.6） % and （95.4 ± 3.2） %, P〈 0.01）. The expression rate of CD14 in the emodin group was （8.4 ± 2.8） %, which was significantly higher than that in the control group （3.7 ± 2.3） % （P 〈 0.01 ）. There were no significant difference between the emodin group and the control group about expression rates of HLA-DR and CD11c （P 〉 0.05）. CONCLUSION Emodin could evidently interfere with the maturation of DC and the expression of costimulatory molecules on surface of DC.