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目的:观察在机械牵张应变作用下,人牙周膜细胞(HPDLCs)内磷脂酶C-γ1(PLC-γ1)水平的变化。方法:通过原代和传代培养,获得性状稳定的HPDLCs,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。加载于不同的时间段,通过流式细胞仪和激光扫描共聚焦显微镜对细胞进行PLC-γ1水平的检测。应用SPSS 13.0软件对数据进行方差分析。结果:使用流式细胞仪对细胞内PLC-γ1水平检测发现: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)。随着加载时间的继续延长,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:对照组和动态加载组都有PLC-γ1的表达,但加载50 min组表达相对较强。结论:机械牵张应变可以引起HPDLCs内PLC-γ1水平的变化。
Objective: To observe the changes of phospholipase C-γ1 (PLC-γ1) level in human periodontal ligament cells (HPDLCs) under mechanical stretch stress. METHODS: HPDLCs with stable traits were obtained by primary and subculture. Cells were loaded with 1%, 10% and 20% dynamic stretch strain using a dynamic mechanical strain cell loader. Loaded at different time periods, and the cells were tested for PLC-γ1 levels by flow cytometry and laser scanning confocal microscopy. Data were analyzed for variance using SPSS 13.0 software. Results: The intracellular PLC-γ1 level was detected by flow cytometry. The level of PLC-γ1 in cells was maintained at a low level during the initial loading period. With the extension of strain-distraction loading time, γ1 levels gradually increased. At 50 min, the level of PLC-γ1 reached the highest value in each strain group (P <0.01). With the extension of loading time, the levels of PLC-γ1 in each strain group decreased at 60 min (P <0.01). Laser scanning confocal microscopy showed that PLC-γ1 was expressed both in the control group and the dynamic loading group, but it was relatively strong in the 50 min loading group. CONCLUSIONS: Mechanical stretch strain can cause changes in PLC-γ1 levels in HPDLCs.