MyD88 RNA干扰抑制小鼠骨髓树突状细胞成熟的实验研究.pdfVIP

陈家军等 MvD88 RNA干扰抑制小鼠骨髓树突状细胞成熟的实验研究 第9期 · 793· MyD88 RNA干扰抑制小鼠骨髓树突状细胞成熟的实验研究① 陈家军 孙宗全 苏 刚 刘 超 刘金平 邓勇志 史嘉玮 (华中科技大学同济医学院附属协和医院心外科，武汉430022) 中国图书分类号 R392．11 文献标识码 A 文章编号 1000-484X(2007)07-0793-05 [摘 要] 目的：针对小鼠骨髓树突状细胞(Dc)髓性分化因子88(MyD88)，采用RNA干扰技术抑制其合成，观察脂多 糖(LPS)刺激后Dc生物学活性的变化，探讨获得耐受性Dc的方法，为Dc的临床应用提供新思路和理论依据。方法：培养小 鼠骨髓源性Dc，分为对照组、LPS组及RNA干扰组，对照组不予任何处理，LPS组加入终浓度为1 g／ml的LPS，RNA干扰组 于加入MyD88 siRNA 12小时后给予1 ml LPS刺激，继续培养3天。免疫细胞化学检测DC MyD88、核因子一KB(NF—KB)的 表达，Western blot检测DC MyD88的表达；流式细胞术检测Dc细胞表面CD80、CD86及MHC—I1分子的变化，ELISA法检测各 组Dc分泌TNF．OL。IFN．^y和IL一12的浓度，混合淋巴细胞培养检测T细胞增殖能力。结果：LPS促进Dc高表达CD80、CD86 及MHC．I1分子，促进Thl型细胞因子释放、MyD88高表达及NF—KB核移位，并诱导T细胞增殖，MyD88 siRNA可阻断LPS的 这些效应。结论：MyD88 siRNA可抑制Dc成熟，产生耐受性Dc，增强同种未成熟Dc的免疫耐受诱导作用，为进一步研究Dc 的临床应用奠定了基础。 [关键词] 树突状细胞；MyD88；RNA干扰；脂多糖 Inhibition effect of MyD88 siRNA to the activation of dendritic cells from murine bone marrow CHENJia．Jun，SUN Zong·Quan，SU Gnng，LIU Chao，LlU fin—Ping，DENG Yong·Z ，SHI Jia—Wei．Depart· ment of Cardiovascular Surgery，Union Hospital，rongii Medical College，Huazhong University ofScience and Tech· nology．Wuhan 43O022， ina [Abstract] 0bjective：To observe the impact of myeloid differentiaion factor 88(MyD88)siRNA on the biological activities of dendritic cells(DCs)cultured from murine bone marrow under stimulation by Lipopolysaccharides(LPS)，and provide novel ideas and basis of DC’S clinical applications．Methods：Mouse DCs were generated from bone marrow cells and were divided into control group， LPS group and RNA interference group．Control group was added nothing，and LPS group was added LPS at the final concentration of 1 m1．MyD88 siRNA was added in RNA interference group，and 12 hours later LPS at the final concentration of 10 ml was add— ed．AU groups were cultured for 3 days thereafter．Immunochemistry was used to detect the concentration of MyD88 and NF—KB．West。 ern blot was used to detect the concentra