【摘要】 【目的】探讨系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)B细胞刺激因子(Blys)基因和蛋白在体外的表达及IL-10对其表达的影响。【方法】梯度密度离心法分离25例系统性红斑狼疮患者和20名女性健康志愿者外周血单个核细胞,分为2组,IL-10(100ng/mL)组和培养基组(仅含RPMI1640培养基),分别于培养0、6、12、24、72h离心收集PBMCs,RT-PCR法检测刺激后0～24h细胞Blys mRNA表达,流式细胞仪和直接免疫荧光法检测72h膜结合型Blys蛋白表达。【结果】①系统性红斑狼疮患者外周血单个核细胞Blys mRNA和蛋白表达体外高于健康对照(P<0.001)。②IL-10显著增强健康对照和系统性红斑狼疮患者外周血单个核细胞Blys mRNA表达,12h作用最强(0.487±0.058 vs 0.251±0.050,P<0.001;0.638±0.084 vs 0.392±0.059,P<0.001)。③IL-10显著增强健康对照和系统性红斑狼疮患者外周血单个核细胞膜结合型Blys蛋白表达(FACs,4.53±0.71 vs 3.24±0.57,P<0.001;5.79±0.91 vs 4.55±0.83,P<0.001)。④直接免疫荧光法检测外周血单个核细胞膜结合型Blys蛋白显示,IL-10刺激后Blys表达增强。【结论】IL-10可上调系统性红斑狼疮患者外周血单个核细胞Blys基因和蛋白表达。更多还原

【Abstract】 [Objective]To determine the expression of membrane-bound B lymphocyte stimulator (Blys) protein and its mRNA in vitro of peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients, and to investigate the role of interleukin-10 (IL-10) on the expression of Blys. [Methods]PBMCs were obtained from 25 SLE patients and 20 healthy volunteers. They were randomized into IL-10 (100 ng/mL) group and media group. PBMCs were collected at 0, 6, 12, and 24 h for Blys mRNA assessment using semi-quantitative reverse transcription-PCR (RT-PCR). PBMCs were also collected at 72 h for membrane-bound Blys protein detection using flow cytometry and direct immunofluorescence. [Results] ①The expression of Blys mRNA and membrane-bound protein in PBMCs was significantly higher in SLE patients than healthy controls in vitro(P< 0.001). ②IL-10 enhanced Blys mRNA expression in PBMCs in both healthy controls and SLE patients, with the greatest effect at 12 h (0.487±0.058 vs 0.251±0.050,P< 0.001;0.638±0.084 vs 0.392±0.059,P< 0.001).③IL-10 also increased the expression of membrane-bound Blys protein in both healthy controls and SLE patients (FACs, mean fluorescence intensity, 4.53±0.71 vs 3.24±0.57,P< 0.001; 5.79±0.91 vs 4.55±0.83,P< 0.001).④Direct immunofluorescence by con-focus micrography also indicated that IL-10 increase the expression of membrane-bound Blys protein. [Conclusion] In vitro, the expression of Blys mRNA and membrane-bound protein was elevated in SLE patients, and IL-10 had positive effects on its expression.更多还原