脂多糖刺激对牙周膜细胞培养上清中RANKL和OPG水平改变的影响  

The production of receptor activator of NF-kB ligand and osteoprotegerin in culture supernatant of human periodontal ligament cells stimulated with E. coli LPS

作　　者：武海春[1] 许尧生 李彬[1] 

机构地区：[1]河北省衡水市第二人民医院口腔科,053000 [2]哈利逊国际和平医院耳鼻喉科

出　　处：《现代口腔医学杂志》2013年第2期102-105,共4页Journal of Modern Stomatology

摘　　要：目的检测脂多糖(LPS)刺激后,体外培养人牙周膜细胞分泌核因子-kB受体活化子配体(RANKL)和骨保护素(OPG)浓度的变化,探讨其对牙槽骨吸收的直接影响。方法组织块法原代培养正常人牙周膜细胞并进行来源鉴定。以10mg/ml LPS刺激第4代细胞,在不同时间段(0h,4h,8h,12h,24h)用酶联免疫吸附实验(ELISA)检测RANKL和OPG在细胞培养上清中的含量,并进行统计学分析。结果 ELISA结果显示正常牙周膜细胞培养上清中可检测到少量RANKL和OPG,LPS刺激后RANKL的表达则随时间延长而逐渐增强,且4h、8h、12h、24h时间点各实验组均与对照组有显著差异(P<0.01)。各实验组之间,8h、12h两时间点有显著差异(P<0.05),其他时间点之间无显著差异(P>0.05);而LPS刺激后不同时间OPG的含量未见显著变化(P>0.05)。未经LPS刺激的牙周膜细胞培养上清中可检测到少量RANKL和OPG,但随时间延长无显著变化。结论体外培养牙周膜细胞在LPS刺激后一定时间内RANKL表达增强,而OPG表达无明显变化。Objective To examine the production of receptor activator of NF-kB ligand （RANKL） and osteoprotegerin （OPG） in the culture supernatant of human periodontal ligament cells after stimulation with E.coli lipopolysaeeharide （LPS）, and to elucidate the role of periodontal ligament cells in alveolar bone resorption. Methods Primary human periodontal ligament cells were cultured and identified immunologically. The fourth passage of the ceils was stimulated with E coll. LPS for Oh, 4h, 8h, 12h, and 24h, enzyme linked immunosorbent assay （ELISA） was used to examine the concentration of secreted RANKL and OPG in the supernatant. Results After stimulation with LPS, OPG concentration in the supernatant exhibited no significant change, but RANKL concentration in the stimulated groups increased gradually from 4h, and peaked at 24h, statistic analysis revealed significant difference for all the stimulated groups when compared with the control groups （P〈0.01）; within stimulated groups, 8h and 12h also revealed a significant difference （P〈0.05）; Groups without LPS stimulation exhibited a low level of RANKL and OPG, and had no significant changes at any of the time points examined. Conclusion Periodontal ligament cells stimulated with LPS expressed an increased level of RANKL, but OPG expression have no significant change.

关 键 词：人牙周膜细胞 脂多糖 骨保护素 核因子-kB受体活化因子配体 ELISA 

分 类 号：R781.4[医药卫生—口腔医学]