静态张应力作用下人牙周膜干细胞核心结合因子Cbfa1表达的变化

摘要

目的 检测静态张应力作用对人牙周膜干细胞核心结合因子Cbfa1表达变化的影响.方法 有限稀释法克隆化培养纯化人牙周膜干细胞(hPDLSCs),应用内外双层套筒式硅胶膜细胞加力装置对人牙周膜干细胞加力,加力时间点为0、3、6、12、24小时.采用原位杂交和实时定量PCR检测各加力时间点Cbfa1 mRNA的表达;蛋白免疫印迹法(Western blot)检测人牙周膜干细胞受力后Cbfa1蛋白的表达.结果 人牙周膜干细胞受到静态张应力作用后3小时、6小时、12小时Cbfa1 mRNA表达均升高(P<0.05),且以加力3 小时最为明显,6小时后开始下降,24小时后基本恢复到不加力时的表达水平.加力3小时Cbfa1蛋白表达未见明显升高,加力6小时后Cbfa1蛋白逐渐升高,一直持续到加力24小时(P<0.05).结论 静态张应力作用下,人牙周膜干细胞核心结合因子Cbfa1在RNA水平和蛋白水平的表达均发生变化,说明Cbfa1在机械力促使人牙周膜干细胞向成骨细胞方向分化的过程中起到了重要作用.%Objective To examine the expression of core biding factor at (Cbfa1) in human periodonlal ligamenl stem cells( hPDLSCs) loaded with slalic Lensile slress. Methods hPDLSCs were subjected Lo slalic Lensile slress for 0,3 , 6,12,24 h. The expression of Cbfal was delecled by in situ hybridization,real-Lime PCR and Weslern blot. Results The expression of Cbfal mRNA in hPDLSCs increased al 3,6 and 12 h. The expression of Cbfal prolein increased over Lime with the slress loading. Conclusion Slalic lensile slress may induce differentiation of hPDLSCs towards osleoblaslic lineage through increasing expression of Cbfal.