TNF-α对牙周膜干细胞和颌骨骨髓间充质干细胞自噬水平的影响

摘要

目的:探讨肿瘤坏死因子-α(tumor necrosis factor α,TNF-α)对牙周膜干细胞(periodontai iigament stem ceiis,PDLSCs)和颌骨骨髓间充质干细胞(jaw bone marrow mesenchymai stem ceiis ,JBMMSCs)两种干细胞自噬水平的影响. 方法:通过胰酶消化法体外分离培养PDLSCs/JBMMSCs,流式细胞仪及实时定量RT-PCR筛选短时间内不导致细胞凋亡的TNF-α的最大浓度,然后与PDLSCs/JBMMSCs共培养,Western biot检测不同时间点两种细胞自噬和凋亡的表达水平. 结果:成功地筛选出TNF-α的最佳浓度为50 ng/mL. 采用该浓度作用于PDLSCs/JBMMSCs,短期作用(24 h)可以激活细胞的自噬水平,凋亡水平下降;如果TNF-α持续作用,细胞的自噬水平反而下降,凋亡增加. 结论:在一定的条件下,TNF-α可以激活PDLSCs/JBMMSCs的自噬水平,免于细胞发生凋亡.%Objective:To evaiuate the effect of TNF-αon the autophagy ievei in periodontai iigament stem ceiis (PDLSCs) and jaw bone marrow mesenchymai stem ceii (JBMMSCs).Methods:The human PDLSCs/JBMMSCs were isoiated and cuitured by the method of trypsmization ,the fiow cytometry and reai-time quantitative PCR screened the concentration of TNF-α( The maximum concentration that didn′t cause the apoptosis of ceiis in a short period ) .Western biot was appiied to estimate the ievei of autophagy and apoptosis at different time points after PDLSCs/JBMMSCs co-cuituring with TNF-α.Results:We have successfuiiy screened the concentration of TNF-α.When co-cuituring with TNF-α,activated autophagy reduced apoptosis of PDLSCs /JBMMSCs in short time,and vice versa in iong time.Conclusions:The infiammatory cytohines TNF-αcouid activate the autophagy ievei , and avoid apoptosis of PDLSCs/JBMMSCs under certain conditions .