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探讨雷帕霉素(Rapamycin,Rapa)对树突状细胞(DC)表面DC-SIGN及其胞内转录因子PU.1基因表达,以及对DC功能的影响。体外培养人外周血单个核细胞来源的DC,结合PU.1基因特异性siRNA转染,给予不同浓度Rapa处理。采用流式细胞术及Western blot检测DC表面DC-SIGN表达变化;细胞迁移试验检测DC迁移能力;混合淋巴细胞反应检测DC刺激T细胞增殖能力。结果显示,Rapa呈剂量依赖性尤以10ng/ml时可下调DC-SIGN表达(P<0.01),同时可抑制DC胞内PU.1基因表达,以及DC迁移及刺激T细胞能力均受到明显抑制(P<0.01)。另发现,经siRNA沉默PU.1基因后,Rapa对DC-SIGN表达基本无影响。本研究表明,Rapa可通过抑制DC-SIGN表达,从而产生对DC迁移、刺激T细胞能力的调抑效应。这一效应可能与Rapa影响PU.1基因转录途径有关。
To investigate the effect of rapamycin (Rapa) on the expression of DC-SIGN and its intracellular transcription factor PU.1 on dendritic cells (DCs) and on the function of DCs. In vitro cultured human peripheral blood mononuclear cell-derived DCs, combined with PU.1 gene-specific siRNA transfection, given different concentrations of Rapa treatment. The changes of DC-SIGN expression on DCs were detected by flow cytometry and Western blot. The migration ability of DC was detected by cell migration assay. The proliferation of DCs stimulated by DCs was detected by mixed lymphocyte reaction. The results showed that Rapa could down-regulate the expression of DC-SIGN (P <0.01) in a dose-dependent manner especially at 10ng / ml, and at the same time, inhibited the expression of intracellular PU.1 gene, the ability of DCs to migrate and the ability of stimulating T cells (P <0.01). In addition, it was found that Rapa had no effect on the expression of DC-SIGN after silencing PU.1 by siRNA. This study shows that Rapa can inhibit the DC-SIGN expression, resulting in DC migration, stimulating T cell capacity downregulation effect. This effect may be related to the effect of Rapa on the transcription of PU.1 gene.