天津医药

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类风湿关节炎滑膜成纤维细胞对T淋巴细胞生物学行为的干预作用

赵文君1,崔爽爽1,邢国胜2,于顺禄2   

  1. 1. 天津市天津医院骨科研究所
    2. 天津医院
  • 收稿日期:2012-07-09 修回日期:2012-09-03 出版日期:2012-11-15 发布日期:2012-11-15
  • 通讯作者: 赵文君

Effects of Rheumatoid Synovial Fibroblasts on Biological Properties of T cells

  • Received:2012-07-09 Revised:2012-09-03 Published:2012-11-15 Online:2012-11-15

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摘要： 目的：探讨体外培养类风湿关节炎（RA）患者滑膜成纤维细胞对T淋巴细胞生物学行为的影响。方法：收集关节置换的RA患者滑膜组织，酶消化法获得滑膜成纤维细胞并扩增培养，选用第三代细胞以相同浓度与Jurkat T细胞株共育培养。MTT测定共育培养后T淋巴细胞的扩增能力，流式细胞仪检测凋亡率，实时荧光定量PCR方法测定T淋巴细胞白细胞介素（IL）-2mRNA、IL-23p19mRNA表达水平，并分别与T淋巴细胞单纯培养组比较。结果：共育培养的T淋巴细胞在RA滑膜成纤维细胞的刺激下，4 d达到快速增殖期，7 d达到增殖平台期，单纯培养的T淋巴细胞7 d至10 d进入快速增殖期，13 d达到增殖平台期。 4 d、7 d共育培养的T淋巴细胞增殖率明显高于单纯培养T淋巴细胞（P<0.001），10 d、13 d共育培养的T淋巴细胞增殖率低于单纯培养T淋巴细胞（P<0.001）。RA滑膜成纤维细胞抑制共育培养的T淋巴细胞的早期凋亡（P<0.005）。RA滑膜成纤维细胞促进T淋巴细胞因子IL-2mRNA、IL-23p19mRNA的表达。结论：RA滑膜成纤维细胞可诱导T淋巴细胞的激活，这可能是RA发病及进展的影响因素之一。

关键词: 关节炎, 类风湿, 滑膜, 成纤维细胞, T淋巴细胞, 共育培养

Abstract: Objective: To investigate the effect of rheumatoid synovial fibroblast on biological properties of T cell. Methods: The sample of synovial tissue was obtained from RA patient during joint replacement surgery. The synovial fibroblasts were extracted by enzyme. The cells of the third passage were co-cultured with Jurkat T cells under the same concentration. MTT method was used to detect the proliferation ability of the co-culture T cells. Flow Cytometer was used to detect the apoptosis of the co-cultured T cells. Real-Time PCR was used to detect the expression of IL-2 mRNA and IL-23p19mRNA. All of the results were compared with the group of T cells alone, respectively. Results: The fast-proliferation phase of the co-cultured T cells had been observed during the previous four days, and reached the proliferation platform at the 7th day, while that of the T cells alone group was observed during the 7th day to the 10th day, and reached the proliferation platform at the 13th day. The proliferation rate was significantly higher in the co-cultured group than that of T cells alone group at the 4th day and the 7th day respectively (P<0.001). The proliferation rate was significantly lower in the co-cultured group than that of T cells alone group at the 10th day and the 13th day respectively (P<0.001). RA synovial fibroblasts co-cultured with T cells inhibited the apoptosis of the T cells (P<0.005). The expression of IL-2mRNA and IL-23p19mRNA in T cells were significantly increased by RA fibroblasts. Conclusion: RA fibroblasts have the potential effect on activating the co-cultured T cells. This may be one of the factors involved in RA pathogenesis and progression.

Key words: arthritis, rheumotoid, synovial membrane, fibroblasts, T cell, co-culture