细胞因子信号转导抑制分子1干扰前后线粒体M2蛋白对树突状细胞功能的影响

检验医学 ›› 2012, Vol. 27 ›› Issue (9): 764-769.

细胞因子信号转导抑制分子1干扰前后线粒体M2蛋白对树突状细胞功能的影响

周运恒¹,马红霞²,曹广亚³,杨再兴⁴,仲人前⁵

1. 武警上海市总队医院检验科
2. 上海市东方医院体检中心
3. 武警上海市总队医院
4. 第二军医大学附属长征医院实验诊断科
5. 长征医院实验诊断科

收稿日期:2012-03-13 修回日期:2012-05-14 出版日期:2012-09-12 发布日期:2012-09-12
通讯作者: 仲人前
基金资助:
国家高技术研究发展计划（863计划）

Study on influence of mitochondrial M2 protein on dendritic cells before and after silencing suppressor of cytokine signaling 1

Received:2012-03-13 Revised:2012-05-14 Online:2012-09-12 Published:2012-09-12

摘要/Abstract

摘要： 目的探讨细胞因子信号转导抑制分子1（suppressor of cytokine signaling，SOCS1）干扰前后，线粒体M2蛋白对外周血来源的树突状细胞（dendritic cell，DC）功能的影响。方法诱导和培养健康人外周血来源的DC，RNA干扰（RNA interference，RNAi）抑制SOCS1的表达，用不同浓度的M2蛋白刺激DC，用流式细胞术 (flow cytometry，FCM )分析DC表型CD83、CD86的表达，用酶联免疫吸附测定法(enzyme linked immunosorbent assay，ELISA)检测DC培养上清液中白细胞介素-10（interleukin-10，IL-10）和IL-12的变化。结果 DC在M2蛋白浓度为70 μg/ml刺激24 h，35 μg/ml 刺激24 h、48 h和72 h时，与对照组相比，CD83和CD86的表达率，IL-10和IL-12的水平均升高，差异有统计学意义（P<0.05）。M2刺激SOCS1干扰后的DC，在不同刺激浓度和作用时间CD83和CD86表达率以及IL-12水平与单纯M2刺激组相比均升高，差异有统计学意义（P<0.05），而IL-10水平在两者之间无显著性差异（P>0.05）。结论 DC在接受高浓度的M2刺激后，其成熟度、抗原递呈和Th1的极化能力增强；抑制SOCS1的表达，M2可进一步促进DC功能的增强，可能导致自身耐受的破坏。

Abstract: Objective To study the effects of M2 protein on peripheral blood mononuclear cells(PBMC)-derived dendritic cell(DC) after silencing SOCS1. Methods: DC were generated from donor-derived PBMC and treated with SOCS-1 siRNA. DC were stimulated by various concentrations of M2 protein at different times CD83 and CD86 of DC was performed by flow cytometry（FCM）.The levels of interleukin-10(IL-10) and IL-12 in culture supernatant of DC were measured by enzyme linked immunosorbent assay (ELISA). Results The expression of CD83 and CD86, the levels of IL-10 and IL-12 of DC under stimulation of M2 protein at 70 μg/ml after 24 h, at 35 μg/ml after 24 and 48 h were all significantly higher than those in the control group（P<0.05）. After silencing SOCS1 by siRNA of DC in the presence of various concentrations of M2 at different times, the expression of CD83 and CD86, the levels of IL-12 were all increased significantly than those in the M2 group（P<0.05）. However, there were not any significant difference of IL-10 levels of DC between the two groups（P>0.05）. Conclusions The results indicated that DC after stimulation of M2 protein at high concentration had increased capacity to activate Th1 subset proliferation, maturation and antigen presentation. The functions was further enhanced after silencing SOCS1 of DC in the presence of M2, which maybe contribute to the broken of self-tolerance.

周运恒马红霞曹广亚杨再兴仲人前. 细胞因子信号转导抑制分子1干扰前后线粒体M2蛋白对树突状细胞功能的影响[J]. 检验医学, 2012, 27(9): 764-769.

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