高迁移率族蛋白B1对牙周膜成纤维细胞表达细胞因子影响的研究

目的观察高迁移率族蛋白B1（HMGB1）对人牙周膜成纤维细胞表达白细胞介素-6（IL-6）、破骨细胞核因子κB受体活化因子（RANKL）、骨保护因子（OPG）的影响，初步探讨HMGB1在牙周疾病的作用。方法采用原代组织块培养法，培养人牙周膜成纤维细胞，用第4~6代的细胞进行实验。分别用10、30、100 ng·mL-1质量浓度的HMGB1孵育牙周膜成纤维细胞24 h后，RT-PCR检测IL-6、RANKL、OPG的mRNA表达；Western blot法检测RANKL、OPG的蛋白表达。均以0 ng·mL-1质量浓度组为对照，所得数据用单因素方差分析处理。结果HMGB1在10、30、100 ng·mL-1质量浓度时，细胞中的RANKL/OPG mRNA的比值增高（P<0.05），100 ng·mL-1质量浓度时细胞中的IL-6 mRNA的表达量增高（P<0.05）。Western blot检测结果显示10 ng·mL-1质量浓度组的RANKL/OPG的比值有明显增高。结论一定浓度的HMGB1可使牙周膜细胞中的RANKL/OPG比值增高，还会诱导炎症因子IL-6 mRNA表达上调。提示HMGB1可能会在牙周炎的发病以及炎症进展中发挥作用。

Effects of high mobility group box 1 in activating periodontal ligament fibroblasts to express cytokine

Objective To investigate the influence of high mobility group box 1（HMGB1） on the expression of interleukin 6（IL-6）, receptor activator of nuclear factor-kappa B ligand（RANKL） and osteoprotegerin（OPG） on periodontal ligament fibroblasts. Methods Human periodontal ligament fibroblasts were stimulated with HMGB1 at concentrations of 10, 30, and 100 ng·mL-1 for 24 h. RT-PCR and Western blot analysis were performed to check mRNA and protein expression of IL-6, RANKL and OPG on the cells. Results The ratio of RANKL/OPG was increased at both mRNA and protein level after HMGB1 stimulation at 10, 30, 100 ng·mL-1. Inflammatory cytokine IL-6 was upregulated by HMGB1 at the concentration of 100 ng·mL-1. Conclusion Increased ratio of RANKL/OPG and IL-6 on periodontal ligament fibroblasts suggests that HMGB1 might play a role in the pathogenesis and progression of periodontal disease.