TNF―α对成骨细胞程序性坏死影响及相关机制研究.docVIP

TNF―α对成骨细胞程序性坏死影响及相关机制研究 　　[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， RIP3 and MLKL were detected by Western blot. Results Compared with the normal control group， the proliferation activity of TNF-α group was decreased and the number of programmed necrosis was increased （P 0.05）. After the Nec-1 treatment， the proliferative activity was significantly increased compared with TNF-α group， and the number of programmed necrosis was decreased （P 0.05）. As time went on， the differences between the groups became more pronounced （P 0.05）. Compared with the normal control group， expression of RIP3 and MLKL protein in TNF-α group was significantly lower than that in normal control （P 0.05）. After the Nec-1 treatment， the expression of RIP3 and MLKL protein had significantly differences compared with TNF-α group （P 0.05）. Conclusion TNF-α can inhibit osteoblast proliferation and promote programmed necrosis of osteoblasts， which may be accomplished b