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作 者:李羽佳[1] 孟祥慧[2] 孙婷婷[1] 吴可非[1] 雷蕾[1] 杨宇[2] 秦岭[2] 杨娉婷[1] 肖卫国[1]
机构地区:[1]中国医科大学附属第一医院风湿免疫科,沈阳110001 [2]中国医科大学基础医学院生理学教研室
出 处:《中华风湿病学杂志》2013年第7期474-476,I0001,共4页Chinese Journal of Rheumatology
基 金:国家自然科学基金(81271324,30801204);辽宁省自然科学基金(201202250);辽宁省医学高峰建设工程重点科研项目(200932,2010006)
摘 要:目的明确CD4+CD25+T细胞对小鼠破骨细胞的形成及骨吸收功能的影响。方法应用免疫磁珠分选方法分离C57BL/6小鼠CD4+CD25+T细胞和CDllb+单核细胞。在M-CSF及RANKL存在下,将分离的CDllb+细胞培养3d,然后与CD3激活的CD4+CD25+T细胞或CD4+CD25-T细胞再共同培养4d。CDllb+细胞单独培养组为对照组。观察各组破骨细胞数目及骨吸收陷窝的数目。统计学方法采用单因素方差分析和U检验。结果所获得的CD4+CD25+T细胞,CD4+CD25-T细胞及CDIlb+细胞纯度均为90%以上。3组破骨细胞的形成数目分别为3.4±0.8、16.2±1.3、16.2±2.7(200x电子显微镜下5个视野的平均值);骨吸收陷窝数目分别是15.3±2.8,47.4±4.8及50.3±5.0(100x电子显微镜下10个视野的平均值)。ANOVA及U检验的结果显示,CD4+CD25+T组的破骨细胞形成数目及骨吸收陷窝数目显著少于其他2组(P〈O.01)。结论CD4+CD25+T细胞对破骨细胞的形成及其骨吸收功能存在抑制作用,为阐明免疫异常所伴发的骨质疏松的机制提供了线索。Objective To reveal the mechanism of inflanunation accompanied osteoporosis by examining the effect of CD4+CD25 + T cells on osteoclasts formation and bone absorption. Methods Mouse CDllb cells and CD4+CD25+ T cells were isolated using the microbead-based kits. The CDllb+ monocytes were cultured alone or cocultured with CD4+CD25+ or CD4+2D25- T cells, respectively. The numbers of osteoclasts and bone absorption pits were compared among the three groups. One-way ANOVA and U test was used for statistical analysis. Results The purities of isolated CD4+CD25+ T cells, CD4+CD25- T cells and CD1lb+ monocytes were all over 90% confirmed by flow cytometry. At day 7, the CD1lb+ monocytes cocul- tured with CD4+CD25 + T cells generated significantly less osteoclasts, bone absorption pits than those cocultured with CD4+CD25 T cells and those cultured alone (all P〈0.01). The number of osteoclasts of the three groups was 3.4±0.8, 16.2±+1.3, 16.2±2.7 (the mean number of osteoblasts of 5 high-power fields with 200 power electronic microscopy) respectively; the number of bone absorption pits was 15.3±2.8, 47.4±4.8 and 50.3±5.0 (the mean number of 10 field with 100 power electronic microscopy)respectively. The number of osteoclasts and bone absorption pits of the CD4+CD25 + T group were significantly lower than the other two groups (all P〈0.01). Conclusion CD4+CD25. T cells inhibit osteoclasts formation and its bone absorption effect, which highlights the role of CD4+CD25+ T cells on osteoporosis due to immune dysfunction.
关 键 词:破骨细胞 骨质疏松 CD4+CD25+T细胞
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