内毒素条件下苯妥英钠对人牙周膜干细胞钙调蛋白表达的影响

Expression of CaM in human periodontal ligament stem cells stimulated by PHT and/or LPS

目的:探讨内毒素(LPS)环境下,苯妥英钠(PHT)对人牙周膜干细胞钙调蛋白(CaM)表达的影响。方法:体外培养人牙周膜干细胞,设空白对照、LPS 5μg/ml、PHT 5μg/ml、PHT 20μg/ml、PHT 5μg/ml+LPS 5μg/ml和PHT 20μg/ml+LPS 5μg/ml共6个实验组,刺激24 h、3 d、5 d后,运用RT-PCR、免疫荧光细胞化学技术结合共聚焦显微镜检测各实验组牙周膜干细胞中CaM表达的变化。结果:①适宜浓度的PHT、LPS单一刺激、PHT和LPS共同刺激均可增强牙周膜干细胞CaM的表达。②PHT 20μg/ml对牙周膜干细胞产生CaM的正向作用最强。③PHT 5μg/ml+LPS 5μg/ml的混合刺激作用强于PHT20μg/m+LPS 5μg/m。结论:PHT可能是通过CaM途径对牙周膜干细胞的增殖产生促进作用。低浓度的PHT和LPS混合刺激作用要优于较高浓度的PHT与LPS的混合刺激。

Objective： To investigate the effect of PHT and/or lipopolysaccharide（LPS） on the mRNA and protein expression of calmodulin（CaM） expression in human periodontal ligament stem cells（hPDLSCs）.Methods： hPDLSCs were cultured and identified in vitro.The cells were treated by LPS or PHT,or in combination at various concentrations.After simulating for 24 h,3 d,5 d,RT-PCR and cell immunofluorescence with confocal laser scanning microscope and fluorescent confocal microscopy were used to detect the expression of CaM mRNA and protein expression in the treated cells respectively.Results： ① PHT and LPS alone or in combination at certain concentration enhanced the expression of CaM in hPDLSCs.② 20 μg/ml PHT showed the strongest positive effect.③ PHT at 5 μg/ml combined with LPS at 5 μg/ml was stronger than PHT at 20 μg/ml plus LPS at 5 μg/ml.Conclusion： PHT may promote PDLC preliferation through the way of increasing CaM expression.