刘凯 卓鸿武 刘建忠

    [摘要] 目的 分析TNF-α对成骨细胞程序性坏死的影响并分析其机制。 方法 分离并常规培养小鼠乳鼠成骨细胞，采用TNF-α干预诱导细胞程序性坏死，Nec-1干预抑制细胞坏死。并采用正常培养的成骨细胞为正常对照组。MTT法检测细胞增殖情况；Tunel和Caspase 3双染法检测细胞的程序性坏死情况；Western blot检测细胞RIP1、RIP3、MLKL蛋白表达水平。 结果 与正常对照组比较，TNF-α组增殖活性降低，程序性坏死数量增加(P < 0.05)；与TNF-α组比较，TNF-α+Nec-1组增殖活性显著增加，程序性坏死数量降低(P < 0.05)；随着时间延长，各组之间的差异更加显著(P < 0.05)。与正常对照组比较，TNF-α组RIP3、MLKL蛋白表达降低(P < 0.05)；TNF-α+Nec-1组RIP3、MLKL蛋白表达高于TNF-α组(P < 0.05)。 结论 TNF-α能够抑制成骨细胞增殖，促进细胞发生程序性坏死，这可能是通过降低成骨细胞的RIP3、MLKL蛋白表达来完成的。

    [关键词] 肿瘤坏死因子α；成骨细胞；程序性坏死

    [中图分类号] R684 [文献标识码] A [文章编号] 1673-7210(2018)03(c)-0018-05

    [Abstract] Objective To explore the effect and its related mechanism of TNF-α on programmed necrosis of osteoblasts. Methods Mice osteoblasts were isolated and cultured from suckling mice， and TNF-α was used to induce programmednecrosis of osteoblasts， Nec-1 was used to inhibit cell necrosis. Normal cultured osteoblasts were used as normal control group， and MTT assay was used to detect the cell proliferation. Tunel and Caspase 3 double staining was used to detect the programmed necrosis of osteoblasts. The expression of RIP1 ......