牵张应变作用下人牙周膜细胞内磷脂酶C-γ1水平变化的实验研究

Effects of mechanical stretching on PLC-γ1 in human periodontal ligament cells

目的观察人牙周膜细胞(HPDLCs)内磷脂酶C-γ1(PLC-γ1)水平在机械牵张应变作用下的变化。方法通过原代和传代培养,获得性状稳定的人牙周膜细胞,使用动态机械应变细胞加载仪对细胞进行1%、10%和20%动态牵张应变加载。并于加载后10 min、20 min、30 min、40 min、50 min和60 min,通过流式细胞仪和激光扫描共聚焦显微镜检测细胞内PLC-γ1水平的变化。应用SPSS13.0软件包对数据进行统计分析。结果流式细胞仪检测发现:加载初期,细胞内的PLC-γ1水平维持在较低水平,随着牵张应变加载时间的延长,牙周膜细胞内PLC-γ1水平逐渐升高。50 min时PLC-γ1水平达到了各个应变组的最高值(P<0.01),60 min时各应变组的PLC-γ1水平都有所下降(P<0.01)。激光扫描共聚焦显微镜检测发现:各组都有PLC-γ1的表达,其中50 min加载组表达相对较强。结论机械牵张应变可以引起人牙周膜细胞内PLC-γ1表达量发生改变。

Purpose The purpose of this study is to investigate the effects of mechanical stretching on concentration of PLC-γ1 in the human periodontal ligament cells （HPDLCs）.Methods Primany human periodontal ligament cells were obtained and passaged.Then the cells were stretched by dynamic mechanical strain unit with 1％,10％ and 20％ strain for different time.PLC-γ1 was examined through Flow cytometry and Laser Scanning Confocal Microscope （LSCM）.The data were analyzed using SPSS 13.0 software package.Results In early period of stretching,The expression of PLC-γ1 remained at a lower level.Along with the stretching time prolonged,it increased gradually.The level of PLC-γ1 of each stretching group reached peak at 50 minute （P＜0.01）,then it declined at 60 minute （P＜0.01）.There was also expression of PLC-γ1 in stretching and control cells by LSCM,but the one in 50 minute stretching cells was more than that of control cells.Conclusion Stretching force induced the change of concentration of the PLC-γ1 of HPDLCs.