葡萄糖对人牙周膜成纤维细胞OPG、RANKL mRNA表达的影响

摘要

Objective To observe the effects of different concentrations of glucose on mRNA expression of osteoprotegerin (OPG) and receptor activator of NF-kappa B ligand (RANKL) in human periodontal ligament fibroblasts (HPDLF) and investigate the effects of glucose on alveolar bone absorption. Methods The primary HPDLF were cultured according to the method of tissue explant in vitro and identified by immunohistochemistry. HPDLF from passages four to eight were used for experiments. HPDLF were exposed respectively to different concentrations of glucose (0, 5.5, 11.1, 16.7,22.2, 33.3mmol/l). The mRNA expression of OPG and RANKL in HPDLF was detected using semiquantitative reverse transcription polymerase chain reaction (RT-PCR) analysis. Results High concentrations glucose down-regulated mRNA levels of OPG and up-regulated the mRNA levels of RANKL dose-dependently. The ratio of RANKL mRNA to OPG mRNA also increased as the glucose concentration rose. Conclusion The high level glucose in periodontal tissue in diabetes mellitus could down -regulate OPG expression and up -regulated RANKL expression, increase the ratio of RANKL to OPG, activate the differentiation of osteoclasts and promote alveolar bone absorption, which might increase the susceptibility of diabetics to periodontal disease.%目的:以体外培养的人牙周膜成纤维细胞(HPDLF)为研究对象,观察不同浓度的葡萄糖对HPDLF中骨保护素(OPG)、核因子kB受体活化剂配体(RANKL)mRNA表达的影响,探讨葡萄糖在牙槽骨吸收中的作用.方法:组织块法体外原代培养HPDLF并鉴定,选择4～8代的HPDLF作为实验的靶细胞,不同浓度的葡萄糖(0、5.5、11.1、16.7、22.2、33.3mmol/1)干预HPDLF,半定量逆转录聚合酶链反应(RT-PCR)检测HPDLF中OPG、RANKL mRNA的表达.结果:高浓度的葡萄糖可下调HPDLF中OPG mRNA的表达,呈剂量依赖性;可上调RANKL mRNA的表达,也呈剂量依赖性,且使RANKL/OPG mRNA的比值随着葡萄糖浓度的增高而呈现持续上升的趋势.结论:糖尿病时,牙周组织中高浓度的葡萄糖,可下调OPG的表达,上调RANKL的表达,使RANKL/OPG的比值升高,调节破骨细胞分化,刺激骨吸收,导致牙槽骨的破坏,加重牙周病的病情.