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  

The influence of mechanical stretch on expression of NLRP3 inflammasome related factors in human periodontal ligament cells

作　　者：赵丹[1,2] 吴雅琴[1,2] 胥春 张富强[1,2] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔修复科,上海200011 [2]上海市口腔医学研究所,上海市口腔医学重点实验室,上海200011

出　　处：《医用生物力学》2016年第3期272-277,共6页Journal of Medical Biomechanics

基　　金：国家自然科学基金资助项目(31470903,31270991,30900282);上海市浦江人才计划资助项目(13PJD021);上海市青年科技启明星计划(A类)项目(10QA1404200);上海市重点学科建设项目(T0202,S30206-sms02)

摘　　要：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 a20% 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.

关 键 词：人牙周膜细胞 机械牵张 炎症体 体外培养 

分 类 号：R318.01[医药卫生—生物医学工程]