【作者】 张岚； 周学东； 王倩； 王瑶； 唐路； 郑庆华； 黄定明；

【Author】 ZhangLan~(1,2),ZhouXuedong~(1,2),WangQian~2,Wang Yao~2,Tang Lu~2,Zheng Qinghua~2,Huang Dingming~(1,2*)(1.State Key Laboratory of Oral Diseases,Sichuan University 2.West China school of stomatology,Sichuan University)

【机构】 口腔疾病研究国家重点实验室(四川大学)； 四川大学华西口腔医学院；

【摘要】 目的探讨人牙周膜成纤维细胞在感染状态下,热刺激对其核因子-κB受体活化因子配体(RANKL)及骨保护素(OPG)表达的影响。材料方法采用体外原代培养的人牙周膜成纤维细胞,分别接种在35mm培养皿中进行实验。实验分为两组:一组细胞直接接受不同温度(39℃,43℃,47℃,50℃)的热刺激5min;另一组细胞预先接受1 0ng/mlE.coli LPS刺激24h,然后再接受不同温度的热刺激(39℃,43℃,47℃,50℃)5min。刺激结束后,将所有细胞转移至37℃孵箱中继续培养6小时,分别采用实时荧光定量PCR技术和E LISA法检测RANKL、OPG基因水平和蛋白水平的变化。结果在细胞接受E.coliLPS预刺激24h的感染状态下行热刺激,RANKL、OPG的基因转录受到抑制,但是仍会在特定条件下诱导蛋白分泌。在低热(39℃)条件下,LPS占主导作用,RANKL分泌升高,OPG分泌下降,RANKL/OPG相对比值上升;当温度升高后(43℃和47℃),热刺激对LPS诱发的炎症反应产生抑制作用,RANKL分泌下降,OPG分泌升高,相对比值降低;当温度继续增高,超过一定限度(50℃)后,RANKL/OPG相对比值再次升高。结论当细胞长期处于慢性低毒的感染环境中时,适当的热刺激不会引起RANKL/OPG相对比值上升,局部组织发生骨吸收的可能性较小,只有在低热或极高热条件下才有可能诱发局部组织发生骨吸收。更多还原

【Abstract】 Aim The aim of the study is to investigate the changes in the expression levels of RANKL and OPG,as well as their relative expression ratio,in periodontal ligament fibroblasts when they are treated with heat in infected conditions.Materials and Methods Periodontal ligament fibroblasts were heated at different temperatures(39℃,43℃, 47℃,50℃)for 5min with or without the pretreatment of 10ng/ml E.Coli LPS,and then maintained at 37℃for 6h.After that,the expression levels of RANKL and OPG were investigated using real-time RT-PCR and ELISA,respectively.As a control,the cells were cultured at 37℃.Results When the cells were stimulated with E.coli LPS 24h ahead of heat treatment,there were significant decreases in the mRNA expression levels of RANKL and OPG.Heat stress induced the protein release of RANKL at 39℃in infected situations,while that of OPG was down-regulated.The relative RANKL/OPG expression ratio was increased dramatically.However,when the temperature rose to 43℃and 47℃, respectively,LPS exerted an inhibitory effect on the release of RANKL,as well as their relative expression ratios.Conclusion These results suggested that heat stress affect the balance between RANKL and OPG in periodontal ligament cells in infected conditions in vitro.更多还原