中间普氏菌培养上清对牙龈成纤维细胞RANKL／OPG表达的影响

摘要

Objective:To evaluate the effects of Prevotella intermedia supernatant on the expression of receptor activator of NF-κB ligand (RANKL)and osteoprotegerin (OPG)in human gingival fibroblasts (HGFs).Methods:The HGFs were treated with gradient Prevotella intermedia supernatant.HGFs were observed under the microscope.The real-time RT-PCR and Western blot analysis were used for testing the expression of RANKL and OPG,and the RANKL/OPG ratio was calculated.Results:The number of cells decreased with increase of the supernatant concentration.At 50 μg/mL,no cell survival could be observed under the microscope.The ratio of RANKL/OPG was increased in the cells treated with 12.5 μg/mL of Prevotella intermedia supernatant(P<0.01).Conclusions:Prevo-tella intermedia supernatant could disrupt the homeostasis of RANKL/OPG,destroy the microenvironment of osteoclast differentiation. Prevotella intermedia supernatant may play a regulatory role in osteoclast differentiation and bone resorption in periodontitis.%目的：探讨中间普氏菌培养上清对牙龈成纤维细胞核因子-κB受体活化因子配体（receptor activator for nuclear factor-κB ligand，RANKL）、骨保护素（osteoprotegerin，OPG）表达的影响。方法：将系列浓度的中间普氏菌培养上清作用于牙龈成纤维细胞，显微镜下观察细胞的变化。实时定量 RT-PCR法和 Western blot法分别检测 RANKL、OPG mRNA及蛋白水平的表达，并计算 RANKL/OPG的比值。结果：牙龈成纤维细胞的数量随中间普氏菌培养上清浓度的增加而减少，50μg/mL 时，显微镜下没有活细胞存在。在mRNA水平上和蛋白水平上，RANKL/OPG的比值在12．5μg/mL处理组均高于对照组，差异具有统计学意义（P＜0．01）。结论：中间普氏菌培养上清可诱导人牙龈成纤维细胞表达 RANKL 和 OPG，破坏 RANKL/OPG比例的平衡，影响破骨细胞分化的细胞因子微环境，在破骨细胞分化和牙周炎骨吸收中起着一定的调节作用。