体外培养破骨细胞TRAF6的表达

目的:观察体外培养破骨细胞过程中肿瘤坏死因子受体相关因子6(TRAF6)的表达,探讨其与破骨细胞功能的相关性。方法:应用1, 25(OH)2D3 体外诱导大鼠骨髓细胞形成破骨细胞,观察记录抗酒石酸酸性磷酸酶(tartrate-resistantacidphosphatase, TRAP)阳性多核巨细胞的数目,并采用免疫组织化学的方法检测TRAF6的表达。结果:应用1, 25(OH)2D3 的实验组可见TRAF6的阳性表达,并随培养时间的延长发生表达强度的变化。结论:TRAF6可能参与了体外培养破骨细胞的成熟与分化。     

破骨细胞相关因子在骨再建过程中表达的定量分析

目的 研究肿瘤坏死因子-α（TNF-α）、核因子-?资B受体激动剂（RANKL）、抗酒石酸酸性磷酸酶（TRAP）在骨再建过程中的表达。方法 用8周龄雄性C57BL/6J小鼠建立骨折模型,用RT-PCR法对骨形成过程中TNF-α、RANKL和TRAP的表达进行时程分析。结果 骨折后TNF-α、RANKL、TRAP的表达均上调, 但峰值在不同的时间点出现。TNF-α在骨折后2 d达峰值, RANKL的峰值出现在骨折5 d后, 而TRAP的表达在骨折后10 d达到峰值。结论 骨的再建也有破骨细胞相关因子的参与, 它有赖于成骨和破骨的精确平衡。     

核因子κB受体活化剂研究进展

核因子κB受体活化剂，是肿瘤坏死因子受体超家族成员，是骨保护素配体在破骨细胞上的受体，骨保护素配体通过与核因子κB受体活化剂结合，将细胞分化等信号传人破骨细胞前体内，促进其分化、融合，并促进成熟破骨细胞的激活过程。     

核因子κB受体活化因子配体和肿瘤坏死因子α经炎性牙周膜干细胞外泌体促进破骨细胞分化

目的　慢性牙周炎表现的病理性骨吸收非常常见,牙周膜干细胞（PDLSCs）的外泌体（Exo）对骨吸收的作用和影响尚不明确,本研究分析了该Exo蛋白组分对破骨细胞分化的影响。方法　分别从正畸前拔牙患者和慢性牙周炎患者的牙周膜组织中提取PDLSCs,通过流式细胞术检测表面标记物;通过差速离心分别提取2种细胞的Exo,即Exo-WT和Exo-CP,并通过Western blot、透射电子显微镜（TEM）、蛋白浓度检测、纳米粒径追踪检测Exo特征;通过蛋白质谱检测2种Exo的蛋白组分;通过酶联免疫吸附（ELISA）验证了差异表达蛋白肿瘤坏死因子α（TNF-α）、核因子κB受体活化因子配体（RANKL）、白细胞介素（IL）-1α、转化生长因子β（TGF-β）、骨形态发生蛋白2（BMP-2）表达水平;将10、100、1 000 μg·mL^-1的Exo-CP或Exo-WT加入RAW264.7培养基中并于5 d时通过实时荧光定量聚合酶链反应（RT-qPCR）、Western blot、抗酒石酸酸性磷酸酶（TRAP）染色检测破骨分化相关指标表达情况;采用SPSS 24.0软件对实验数据进行统计学分析。结果　Exo-CP富集的差异表达蛋白主要与破骨细胞分化的TNF信号通路、核转录因子κB（NF-κB）信号通路相关;ELISA实验证实了Exo-CP中高表达TNF-α、RANKL、IL-1α,低表达TGF-β1、BMP-2（P<0.05）;Exo-CP作用于RAW264.7,显著提高了细胞的破骨分化相关基因及蛋白的表达水平,TRAP染色可见分化的破骨细胞,且呈现浓度依赖性,100、1 000 μg·mL^-1浓度Exo-CP对破骨细胞分化的促进作用显著高于10 μg·mL^-1浓度组（P<0.05）。结论　慢性牙周炎的病理性骨吸收可能由炎性PDLSCs所分泌的Exo通过促进破骨细胞分化引起,Exo中主要的作用蛋白可能为RANKL和TNF-α,本研究为慢性牙周炎骨吸收的发病机制提供了新视角。     

核因子-κB受体活化因子配体在大鼠正畸牙压力侧牙周组织中的表达

目的：探讨正畸牙移动中，压力侧牙周组织中核因子-κB受体活化因子配体(RANKL)的表达在破骨细胞诱导和骨改建中的作用。方法：建立大鼠正畸牙移动模型，对压力侧牙周组织中RANKL蛋白的表达及破骨细胞的生成进行检测。结果：TRAP( )破骨细胞在牙移动第3、5和7天时数量明显增多；免疫组化检测显示牙周成骨细胞、骨衬里细胞、牙周纤维细胞和位于骨陷窝内的破骨细胞，是表达RANKL的主要细胞。与对照组RANKL持续弱阳性表达比较，实验组RANKL在正畸牙移动第3、5和7天时出现强阳性表达，其表达变化与破骨细胞的数量变化较为一致。结论：在大鼠正畸牙移动中，RANKL可能在压力侧牙周组织破骨细胞的形成和骨改建中发挥了重要作用。     

肿瘤坏死因子受体相关因子联合核因子-κB激酶结合激酶1和干扰素刺激因子基因在介导DNA疫苗免疫中的作用

肿瘤坏死因子受体相关因子联合核因子-κB激酶结合激酶1(TBK1)基因不但在DNA疫苗诱导体液免疫和细胞免疫等特异性免疫时发挥着重要的作用,而且在诱导固有免疫方面也有其独特的作用。干扰素刺激因子基因(STING)是由TBK1基因诱导Ⅰ型干扰素产生从而在固有免疫反应时不可缺少的组分。本文就TBK1和STING基因在介导DNA疫苗免疫中的作用作一综述。     

牙齿萌出过程中 RANKL mRNA 的表达与破骨细胞的关系

目的:探讨核因子κB受体活化剂配体 RANKL 在大鼠下颌第一磨牙牙胚冠方组织中 mRNA 的表达及破骨细胞的分化情况.方法:运用原位杂交法检测大鼠下颌第一磨牙牙胚冠方组织中 RANKL mRNA 的表达;TRAP 染色观察破骨细胞分化情况.结果:出生1、3、5、7 d大鼠下颌第一磨牙牙胚冠方牙囊成纤维细胞、成釉细胞 RANKL mRNA的阳性表达强于对照组牙龈成纤维细胞(p<0.01).大鼠下颌第一磨牙牙胚冠方出生后1 d破骨细胞多为单核,3 d时多核破骨细胞增多.结论:RANKL mRNA 在大鼠出生后1、3、5、7 d下颌第一磨牙牙囊成纤维细胞、成釉细胞中有表达,可能通过促进破骨细胞的分化及成熟参与牙齿的萌出.     

肿瘤坏死因子受体相关分子6在成牙本质细胞中表达的研究

目的 :研究肿瘤坏死因子受体相关分子 6(TRAF6)是否在成牙本质细胞中表达。方法 :通过westernblot、免疫组化染色研究TRAF6蛋白在成牙本质样细胞系MDPC 2 3中的表达。结果 :Westernblot结果显示MD PC 2 3细胞总蛋白中有大小约 60Kda的TRAF6蛋白条带 ;免疫组化法表明TRAF6在MDPC 2 3细胞浆呈阳性表达。结论 :本实验从蛋白水平证实了成牙本质样细胞MDPC 2 3表达TRAF6,提示TRAF6可能是一种新发现的影响成牙本质细胞分化的胞内信号转导分子     

骨保护素/核因子-κB受体活化因子/核因子κB-受体活化因子配体信号分子调控牙萌出的研究进展

牙萌出是牙胚、牙槽骨以及多种类细胞系及多条通路信号分子相互作用共同调控的、复杂有序的生理过程。牙齿萌出需要在牙胚的方形成萌出通道,穿过牙槽骨和口腔黏膜到达功能位置。这一过程主要由破骨细胞直接执行,而骨保护素/核因子-κB受体活化因子/核因子κB-受体活化因子配体信号分子(OPG/RANK/RANKL)信号分子则可通过调节破骨细胞的分化与成熟来调控牙槽骨的改建,以保证牙萌出时方牙槽骨能被正常吸收。牙萌出的具体调控机制尚不明确,该文就OPG/RANK/RANKL信号分子调控牙萌出的研究现状作一综述。     

核因子-κB受体活化因子配体在大鼠正畸牙压力侧骨改建中的作用

目的：初步探讨在正畸牙移动压力侧核因子-κB受体活化因子配基(receptor activator of nuclearfactor-κB ligand，RANKL)的表达在破骨细胞诱导和骨改建中的调节作用。方法：建立大鼠正畸牙移动模型，利用免疫组化的方法对压力侧RANKL的表达及其变化进行检测；并进一步观察了RANKL对大鼠骨髓破骨细胞形成的影响。结果：正畸牙移动压力侧组化结果显示，RANKL阳性表达位于牙周膜细胞和位于骨陷窝内的破骨细胞中，在正畸牙移动第3、5和7天时呈强阳性表达。体外破骨细胞诱导实验结果显示，在巨噬细胞集落刺激因子(macrophage clone stimulating factor，M-CSF)协同作用下，RANKL呈剂量依赖型诱导TRAP阳性细胞生成。结论：大鼠正畸牙移动中，RANKL在压力侧的表达上调有诱导破骨细胞形成的作用，并导致牙槽骨吸收。     

Tumor necrosis factor modulates fibroblast apoptosis, PMN recruitment, and osteoclast formation in response to P. gingivalis infection.

P. gingivalis is an important oral pathogen, which has been closely linked to periodontal disease as well as lesions of endodontic origin. Both infections are associated with a decrease in fibroblast numbers, formation of an inflammatory infiltrate, and bone resorption. The goal of this study was to investigate the role that the host response plays in the capacity of P. gingivalis to stimulate fibroblast apoptosis, PMN recruitment, and osteoclastogenesis. This was accomplished by the use of an in vivo calvarial model in mice with targeted deletion of TNF receptors p55 and p75 and matched wild-type mice. The results indicate that P. gingivalis induces fibroblast apoptosis in vivo and establish for the first time that this involves the stimulation of a host response. Moreover, bacteria-stimulated PMN recruitment and osteoclastogenesis were also dependent upon the host response. The results suggest that much of the damage caused by P. gingivalis infection, including fibroblast apoptosis, at least under some circumstances, results from stimulation of the host response rather than the direct effect of bacterial products. Furthermore, this may represent a more general mechanism by which bacterial challenge induces apoptosis of matrix-producing cells through the induction of TNF.     

Immunohistochemical localization of growth factors fibroblast growth factor-1 and fibroblast growth factor-2 and receptors fibroblast growth factor receptor-2 and fibroblast growth factor receptor-3 in normal oral epithelium,epithelial dysplasias,and squamous cell carcinoma

OBJECTIVES: Fibroblast growth factors (FGFs) and their receptors (FGFRs) have been identified in a variety of carcinomas, but there are few studies concerning their presence in oral cancers. The objective of this study was to determine whether FGF-1, FGF-2, and high affinity receptors FGFR2 and FGFR3 are present in the pathogenesis of oral epithelial dysplasias and oral squamous cell carcinoma. STUDY DESIGN: Sections from formalin-fixed, paraffin-embedded samples of oral normal mucosa (n = 14), epithelial dysplasia (n = 20), carcinoma in situ (n = 10), and squamous cell carcinoma (n = 12) were tested for cytoplasmic staining by standard in situ immunohistochemistry with antibodies for FGF-1, FGF-2, FGFR2, and FGFR3. RESULTS: Staining for FGF-1 is decreased or lost in the development of epithelial dysplasia and carcinoma. Staining for FGF-2 showed increased intensity (although not statistically significant) in oral epithelial dysplasias and squamous cell carcinomas and showed a significant increased expression in the upper layers of dysplasias and stratum spinosum-like cells in squamous cell carcinomas. Staining for FGFR2 showed a statistically significant increase in intensity in all layers of epithelial dysplasias and squamous cell carcinomas. Staining for FGFR3 was found in the upper stratum spinosum cells of normal and dysplastic epithelium and well-differentiated squamous cells in squamous cell carcinomas, with a statistically significant increase in staining intensity in dysplastic and carcinomatous tissues. CONCLUSIONS: The loss of FGF-1 is consistent with loss of differentiation in dysplasias and some squamous cell carcinomas. Changes in the localization of FGF-2 and FGFR2 into upper epithelial layers with increasing dysplasia suggest increased mitotic potential of high level cells. The co-localization of FGF-2 and its high affinity receptors in neoplastic tissues suggests an autocrine mechanism of influence on carcinogenesis.     

Tumor necrosis factor (TNF) inhibitor therapy for rheumatoid arthritis

Rheumatoid arthritis (RA) is a systemic autoimmune disorder characterized by inflammation involving large and small joints. Systemic manifestations as well as involvement of paraoral tissues contribute to morbidity. Tumor necrosis factor (TNF) plays a central role in RA by amplifying inflammation in multiple pathways that lead to joint destruction. Tumor necrosis factor inhibitors were first licensed for clinical use in 1998; 3 have been approved for the treatment of RA: Iinfliximab, etanercept, and adalimumab. The purpose of this paper is to review the pathogenesis of RA, the state of the art of therapy, and the most current information on the safety and efficacy of TNF inhibitors for treatment of RA.     

肿瘤坏死因子和脂多糖影响牙周膜细胞表达骨保护因子的研究

目的　研究脂多糖 (lipopolysaccharide,LPS)和肿瘤坏死因子α(tumornecrosisfactor α,TNF α)对牙周膜细胞增殖及分泌骨保护因子 (osteoprotegerin ,OPG)的影响。 方法　培养牙周膜细胞并形成单细胞克隆 ,培养基中加入LPS和TNF α ,MTT法检测牙周膜细胞增殖水平 ,SandwitchELISA法测定培养上清液中OPG含量。结果　TNF α浓度在 1 0 μg/L以上时可增加细胞对OPG的表达量 (P <0 0 5) ,但由于TNF α同时抑制牙周膜细胞增殖 (P <0 0 5 ) ,因此培养上清中OPG总量无明显变化 (P >0 0 5) ;LPS对牙周膜细胞增殖和OPG表达均无明显影响 ,与TNF α也没有交互作用 (P >0 0 5)。结论　TNF α能刺激牙周膜细胞OPG表达水平的提高 ;牙周膜细胞可能不是牙周炎时LPS直接细胞毒作用的效应细胞     

Tumor necrosis factor mediates temporomandibular joint bone tissue resorption in rheumatoid arthritis

Abstract

Objective. To investigate if TNF, IL-1 or their endogenous controls, in relation to ACPA, are associated with radiological signs of ongoing temporomandibular joint (TMJ) bone tissue resorption and disc displacement in RA patients. Methods. Twenty-two consecutive outpatients with TMJ of RA were included. Systemic inflammatory activity was assessed by DAS28. The number of painful regions in the body and ESR, CRP, RF and ACPA were analyzed. TMJ synovial fluid and blood samples were obtained and analyzed for TNF, TNFsRII, IL-1ra, IL-1sRII and ACPA. The ratios between the mediators and their endogenous control receptors were used in the statistical analysis. Magnetic resonance imaging was performed in closed- and open-mouth positions and evaluated regarding disc position and presence of condylar and temporal erosions of the TMJ. Results. A high TNF level in relation to TNFsRII in TMJ synovial fluid correlated to the degree of TMJ condylar erosion. A high IL-1ra level in relation to TNF in TMJ synovial fluid was also correlated to the degree of TMJ condylar erosion. The total degree of TMJ condylar erosion was correlated with the number of painful regions. Conclusion. This study indicates that TNF in TMJ synovial fluid mediates TMJ cartilage and bone tissue resorption in RA. The study also suggests that the degree of endogenous cytokine control is of importance for development of bone tissue destruction.     

TRAF6在大鼠牙胚发育过程中的表达

目的 :研究肿瘤坏死因子受体相关因子 6(TRAF6)在大鼠牙胚发育过程中的表达并探讨其意义。方法 :制备大鼠牙胚发育各阶段标本 ,进行TRAF6的免疫组化研究。结果 :TRAF6在牙胚发育中呈动态时空表达。结论 :TRAF6可能是新发现的一种参与调控牙胚细胞的增殖分化和牙齿发育矿化的胞内信号转导分子。     

Tumor necrosis factor identified in periapical tissue exudates of teeth with apical periodontitis

Root canal samples, taken from periapical tissue exudates during routine root canal treatment procedures, were processed for identification of tumor necrosis factor using a mouse anti-human monoclonal antibody and enzyme-linked immunosorbent assay. Detectable levels of tumor necrosis factor were identified in periapical tissue exudates in chronic apical periodontitis.     

Osteoprotegerin and osteoclast differentiation factor in tooth eruption

A critical cellular event in tooth eruption is the formation of osteoclasts that are needed for bone resorption to form an eruption pathway. To analyze molecular regulation of osteoclast formation and activation, we examined the expression of osteoprotegerin (OPG), an inhibitor of osteoclast formation. In vivo, the gene expression of OPG is reduced in the dental follicle of the first mandibular molar of the rat at day 3 post-natally and in the mouse at day 5. This correlates with the days of maximal mononuclear cell influx and osteoclast numbers in the rat and mouse. Thus, inhibition of OPG gene expression on these days might allow osteoclasts to be formed and/or activated. In vitro studies demonstrated that both colony-stimulating factor-1 and parathyroid hormone-related protein reduced OPG gene expression in follicle cells, suggesting that these are candidate molecules for the in vivo inhibition of OPG expression. Osteoclast differentiation factor (ODF) immunolocalizes to the alveolar bone stromal cells adjacent to the follicle, whereby it might act to stimulate fusion of the mononuclear cells in the follicle.     

Salivary interleukin-6 and tumor necrosis factor alpha in patients with drug-induced xerostomia

It is well known that cytokines are involved in the homeostasis of oral cavity and that altered levels of either serum and/or salivary cytokines have been found in certain oral/systemic diseases. So far, cytokines in connection with xerostomia have been investigated in patients with Sjögren's syndrome. We wanted to find out whether drugs themselves influence salivary glands, which would result in altered cytokine level or whether xerostomia itself of different causes leads to the changes in salivary cytokine levels. Therefore, the aim of this study was to evaluate levels of salivary interleukin‐6 (IL‐6) and tumor necrosis factor (TNF)‐α in 30 patients with drug‐induced xerostomia, age range 29–84 and mean 63.9 years. Control group consisted of 30 healthy participants, age range 30–82 years and mean age 65.2 years. Enzyme‐linked immunoassay was performed on commercially available kits. Statistical analysis was performed by use of Student's test. No significant differences in salivary IL‐6 and TNF‐α between patients with drug‐induced xerostomia when compared with the healthy controls were found (P < 0.05). We might conclude that drugs do not induce damage to the salivary glands which could be seen in altered salivary IL‐6 and TNF‐α levels and that xerostomia itself, induced by drugs does not alter levels of the investigated salivary cytokines.