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。
Objective To explore the effect and its related mechanism of TNF-αon programmed necrosis of osteoblasts.Methods Mice osteoblasts were isolated and cultured from suckling mice,and TNF-αwas 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-αgroup 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-αgroup,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-αgroup 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-αgroup(P<0.05).Conclusion TNF-αcan inhibit osteoblast proliferation and promote programmed necrosis of osteoblasts,which may be accomplished by reducing the expression of RIP3 and MLKL proteins in osteoblasts.
作者
刘凯
卓鸿武
刘建忠
柯典闪
LIU Kai;ZHUO Hongwu;LIU Jianzhong;KE Dianshan(Department of Orthopedics,Fuzhou Second Hospital of Xiamen University,Fujian Province,Fuzhou 350007,China;Department of Geriatrics Rehabilitation,Fujian Institute of Rehabilitation Industry Affiliated Rehabilitation Hospital of Fujian University of Traditional Chinese Medicine,Fujian Province,Fuzhou 350001,China)
出处
《中国医药导报》
CAS
2018年第9期18-21,40,共5页
China Medical Herald
基金
福建省科技厅科技平台建设项目(2015Y2001)
福建省福州市科技计划项目(2017-S-130-6)
