【作者】 黄文雪； 詹雅琳； 郑云飞； 刘娟； 胡文杰； 侯建霞；

【Author】 Wenxue Huang;Yalin Zhan;Yunfei Zheng;Juan Liu;Wenjie Hu;Jianxia Hou;Department of Periodontology,Peking University School and Hospital of Stomatology;Department of Orthodontics,Peking University School and Hospital of Stomatology;

【机构】 北京大学口腔医院牙周科； 北京大学口腔医院正畸科；

【摘要】 目的:牙周膜是维持牙齿稳定的重要结构之一。用全长cDNA文库分析人牙周膜组织中功能活跃基因时发现铁蛋白表达丰富。因此,本研究的目的是探究牙周组织炎症状态下铁蛋白的表达变化,以及铁蛋白对牙周膜细胞的影响。方法:PCR和细胞免疫荧光法检测原代细胞中铁蛋白的表达。丝线结扎法构建小鼠实验性牙周炎模型。免疫组化法检测小鼠牙周组织和人牙龈组织中FTH、FTL、COX2的表达。P.gLPS (0,100,250,500,1000,2000 ng/ml)刺激人牙周膜细胞后,实时定量PCR、western blot实验检测FTH和FTL的表达变化。去铁铁蛋白(0,1,10,100,500,1000 ng/ml)、细胞内通路抑制剂和转铁蛋白受体的小干扰RNA作用于人牙周膜细胞后,实时定量RCP和酶联免疫吸附实验检测IL-6,IL-8,TNF-a基因和蛋白水平表达变化。结果:FTH和FTL在人牙周组织中显著表达并广泛分布。FTH和FTL在炎症性牙周组织中的表达升高,并与炎细胞浸润范围和COX2表达位置相一致。LPS以浓度依赖性和时间依赖性促进牙周膜细胞中FTH和FTL的表达。不同浓度的去铁铁蛋白通过转铁蛋白受体激活ERK/JNK/P38通路,促进牙周膜细胞表达IL-6,IL-8,TNF-a。结论:铁蛋白在人牙周组织中显著表达并广泛分布,在炎症状态下表达升高,并促进牙周膜细胞中炎症因子的表达。铁蛋白在牙周炎的发生发展中发挥重要作用。更多还原

【Abstract】 Objective:The periodontal ligament(PDL) is one of the most important tissues for maintaining the homeostasis of tooth.Previous full length cDNA library analyses about active genes in the PDL revealed aboundant expression of ferritin.So the aim of this study is to explore how ferritin is regulated by local inflammation and to investigate the effect of ferritin inPDLCs.Methods:We examined the expression of ferritin in primary cultured cells by polymerase chain reaction(PCR) andimmunocytofluorescence.Experimental periodontitis was induced by ligation of mandibular second molars in mouse.The expression of ferritin light polypeptide(FTL),ferritin heavy polypeptide(FTH) and COX2 wereexamined by immunohistochemical staining in mouse PDL tissues and human gingiva tissues in vivo.After P.gingivalisLPS(0,100,250,500,1000,2000 ng/ml) stimulatedhPDLCs in vitro,the expressions of FTHand FTL were investigated using real-time PCR and western blot.Then,mRNA and protein levels of IL-6,IL-8 and TNF-a were measured using real-time PCR and enzyme-linked immunosorbent assay(ELISA),after stimulation with iron-free ferritin(0,1,10,100,500,1000 ng/ml),knockingdown transferrin receptor(CD71) by small interfering RNA and incubating with multiple intracellular signaling pathway inhibitors,respectively.Results:FTH and FTL wereconstitutively expressed and widely distributed in periodontal tissues of human.FTH and FTL increased substantially in inflammatory periodontal tissues,which correlated well with the extent of inflammatory infiltration and the expression of COX2.LPS promoted the expressions of FTH and FTL in the manner of concentration-dependence and time-dependence.Apoferritin(iron free ferritin) induced the expression of IL-6,IL-8 and TNF-a in hPDLCs through activating ERK/JNK/P38 signaling pathways and binding with CD71 at various concentrations.Conclusion:Ferritin is constitutively expressed and widely distributed in periodontal tissues of human.Ferritin is upregulated by inflammation and promotes pro-inflammatory cytokine expression in PDL cells.ferritin plays an important role in the etiology and process of periodontitis.更多还原