牵张应变对人牙周膜细胞中NLRP3炎症体通路相关因子表达的影响

目的 研究牵张应变对人牙周膜细胞（human periodontal ligament cells, HPDLCs）表达炎症体通路相关因子的影响。方法 对体外培养的HPDLCs施加20%牵张应变6、24 h，分别采用实时定量PCR以及免疫印迹技术检测NLRP3、Caspase-1及IL-1β的mRNA及蛋白表达的变化。对体外培养的HPDLCs施加20%牵张应变1、2、4、6 h后，采用酶联免疫吸附实验检测细胞分泌的IL-1β蛋白变化。对照组HPDLCs培养于弹性培养皿中，与动态牵张应变加载组的培养条件一致，但不加载应变。结果 6 h牵张应变作用下HPDLCs中NLRP3、 caspase-1及IL-1β的mRNA及蛋白表达较对照组增加（P<0.05）；24 h牵张应变作用下HPDLCs中NLRP3的蛋白表达水平上调（P<0.05）；加载牵张应变4、6 h后，细胞培养上清中IL-1β含量随牵张时间的增长而增加（P<0.05）。结论 20%机械牵张6 h能诱导HPDLCs中NLRP3/Caspase-1炎症体-IL-1β通路因子的表达。
Objective To study the effect of mechanical stretch on expression of inflammasome related factors in human periodontal ligament cells (HPDLCs). Methods HPDLCs were subjected to mechanical stretch with a 20% elongation magnitude for 6 h or 24 h, respectively. The mRNA and protein expression levels of IL-1β, Caspase-1 and NLRP3 were detected by real-time quantitative PCR and Western blot. The levels of IL-1β in the cell-culture medium of HPLCs in response to mechanical stretch for 1, 2, 4, 6 h were detected by ELISA, respectively. In control group, HPDLCs were cultured in similar conditions but not subjected with stretch. Results The mRNA and protein expression levels of IL-1β, Caspase-1 and NLRP3 were up-regulated with mechanical stretch for 6 h (P<0.05). The protein expression level of NLRP3 was up-regulated with mechanical stretch for 24 h (P<0.05). Compared with control group, the content of IL-1β in the cell-culture medium of HPLCs was increased significantly in response to mechanical stretch for 4 h and 6 h(P<0.05). Conclusions The expression of NLRP3/Caspase-1/IL-1β related factors in HPDLCs can be induced by 20% mechanical stretch for 6 h.