牙龈卟啉单胞菌脂多糖影响单核细胞表达Th17细胞分化相关因子的研究  

Effect of lipopolysaccharide of porphyromonas gingivalis on the expression of Th17 cell differentiation related cytokines in monocytes

作　　者：张莉平[1] 高雳[2] 赵雅静[1] 张弛[1] 王兴羽[1] 赵川江[3] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.广东省口腔重点实验室,硕士生广东510055 [2]中山大学光华口腔医学院.附属口腔医院.牙周科,副教授广东510055 [3]中山大学光华口腔医学院.附属口腔医院牙周科,副教授广东510055

出　　处：《中华老年口腔医学杂志》2015年第1期1-6,33,共7页Chinese Journal of Geriatric Dentistry

基　　金：国家自然科学基金(项目编号:30801296);国家自然科学基金(项目编号:81300887)

摘　　要：目的:探讨牙龈卟啉单胞菌脂多糖对人外周血CD14+单核细胞表达Th17细胞分化相关细胞因子IL-1β、IL-6、IL-23和TGF-β的影响。方法:采用脂多糖提取试剂盒提取牙龈卟啉单胞菌标准株ATCC33277和高毒力株W83脂多糖;免疫磁珠法分选健康志愿者外周血CD14+单核细胞。以大肠杆菌脂多糖为对照,1μg/ml牙龈卟啉单胞菌脂多糖处理单核细胞,分别培养12、24、36h后收集细胞和上清液,实时定量PCR及ELISA法检测IL-1β、IL-6、IL-23、TGF-βm R NA和蛋白水平的表达。结果:各型脂多糖均可显著上调单核细胞四种细胞因子m RNA及蛋白水平的表达;上调作用具有时间效应,在12h达到高峰。并且三种菌株脂多糖对不同细胞因子表达的影响存在一定差异。结论:牙龈卟啉单胞菌脂多糖可显著上调CD14+单核细胞表达IL-1β,IL-6,IL-23,TGF-β,可能与牙周炎症环境中Th 17细胞的分化相关。Objective：To investigate the effects of lipopolysaccharide（LPS） from Porphyromonas gingivalis（Pg） on the expression of Th17 cell differentiation related cytokines including IL-1β, IL-6, IL-23 and TGF-β in CD14＋ monocytes.Methods： The LPS was extracted from standard strain ATCC33277 and high virulent strain W83 by LPS Extraction Kit.The CD14＋monocytes were separated from human fresh peripheral blood by Anti-Human CD14 Magnetic Particles – DM,and then treated with LPS from Pg as well as Escherichia coli（E.coli） in 1μg/ml for 12、24、36 hours. The expression of IL-1β, IL-6, IL-23 and TGF-β m RNA in CD14＋ monocytes was detected by Real Time- Polymerase Chain Reaction（RT-PCR）. And the protein levels of these cytokines in cell culture supernatant were measured by enzyme-linked immunosorbent assay（ELISA）. Results： Pg-LPS and Ecoli-LPS could up-regulate the expression of IL-1β, IL-6, IL-23 and TGF-β both in m RNA and protein levels in a time-effect manner with peak values at 12 h. And there were differences in the expression of the four cytokines among different LPS treated groups. Conclusion： Pg-LPS influenced the expression of Th17 cell differentiation related cytokines in CD14＋monocytes, which may be associated with the Th17 cell differentiation in periodontal inflammatory context.

关 键 词：牙龈卟啉单胞菌 脂多糖 TH17细胞 细胞因子 单核细胞 

分 类 号：R780.2[医药卫生—口腔医学]