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Investigation of the molecular mechanisms through which osteoclast differentiation regulating factors modulates osteoarthritis progression.

研究破骨细胞分化调节因子调节骨关节炎进展的分子机制。

基本信息

批准号：
21591935
负责人：
KOGA Daisuke
金额：
$ 2.91万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2011
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21591935/
关键词：
OPG RANK RANKL 老化関節軟骨 TNF

项目摘要

PurposeProinflammatory cytokines, such as IL-1 and tumor necrosis factorα(TNFα), are suspected of causing damage to osteoarthritis(OA) cartilage. TNFαcontent is elevated in the synovial fluid of OA joints. There are 2 cell surface receptors for TNFαincluding p55(TNFRI) and p75(TNFRII). TNFαand TNFRs I and II are up-regulated in OA cartilage. For bone metabolism, mice null for TNFR1 have significantly increased peak bone mass, resulting from elevated bone formation. In vitro, TNFαinhibited mineralization of osteoblasts through regulation of NF-kB activity. Thus, whereas TNFR1 plays roles in osteoblast differentiation, its functions in chondrocyte metabolism is unknown. Endochondral ossification is an essential process for development of OA, which is characterized by cartilage degradation and osteophyte formation. The aim of this study is to investigate the role of TNFR1 in the maintenance of articular tissues.Materials and methodsHistological evaluation of aged TNFR1-/-miceKnee joints o … More f TNFR1-/-and their wild-type(WT) littermates(52-60 weeks of age) were evaluated immunohistologicaly. Whole knee joints were removed by dissection, fixed in 4% paraformaldehyde, and decalcified in EDTA. After dehydration and paraffin embedding, serial 5-□m sagittal sections were made from the whole medial compartment of the knee joint. Three sections were picked up at 100-□m intervals from the weight-bearing region of each medial plateau of tibia, and were stained with Safranin O-fast green and HE. Osteophyte area and articular cartilage thickness were quantified in each slide using Image-Pro Plus 4. 1 software.Surgical induction of OA in TNFR1-/-mice.TNFR1-/-and their wild-type(WT) littermates were surgically induced to develop OA by medial collateral ligament transection and medial meniscectomy. Four weeks after surgery, the mice were euthanized.ResultsAt 52-60 weeks of age, whereas osteophyte formation was detected at the medio-anterior edge of tibial plateau regardless of genotype, osteophyte area was significantly increased in TNFR1-/-mice. Moreover, articular cartilage thickness was reduced at medial end region of tibia plateau in TNFR1-/-mice, suggesting ossification of articular chondrocyte was advanced by TNFR1 deficiency. We next compared osteoarthritis development between adult littermates of wild-type and TNFR1-/-mice by creating a surgical osteoarthritis model through induction of instability to the knee joints. Histological evaluation confirmed that the TNFR1 deficiency caused significant acceleration for osteophyte formation. Degree of cartilage destruction was almost comparable between TNFR1-/-and WT. Real time PCR analysis for epiphyseal chondrocyte from new born mice revealed mRNA expression of type X collagen, a marker for chondrocyte hypertrophy, was significantly elevated in TNFR1-/-mice compared to WT littermate. Currently, we are picking up a number of candidate genes for targets of TNFR1 in the course of chondrocyte hypertrophy using cDNA microarray analysis.ConclusionEndogenous TNFR1 had a protective effect against osteophyte formation through inhibition of type X collagen synthesis. Less

目的：促炎细胞因子IL-1和肿瘤坏死因子α（TNFα）被认为与骨关节炎（OA）软骨损伤有关。OA关节滑液中TNFα含量升高。TNFα有2种细胞表面受体，包括p55（TNFRI）和p75（TNFRII）。TNFα和TNFRs I和II在OA软骨中上调。对于骨代谢，TNFR 1缺失的小鼠具有显著增加的峰值骨量，这是由于骨形成增加。在体外，TNFα通过调节NF-κ B活性抑制成骨细胞矿化。因此，尽管TNFR 1在成骨细胞分化中起作用，但其在软骨细胞代谢中的功能尚不清楚。软骨内骨化是OA发生发展的重要过程，其特征是软骨降解和骨赘形成。本研究的目的是探讨TNFR 1在维持关节组织中的作用。材料和方法老年TNFR 1-/-小鼠膝关节的组织学评价。 ...更多信息 f用免疫组织学方法评价TNFR 1-/-及其野生型（WT）同窝仔（52-60周龄）。通过解剖取出整个膝关节，在4%多聚甲醛中固定，并在EDTA中脱钙。经脱水、石蜡包埋后，取膝关节内侧间室全层连续5-□m矢状切片。从每个胫骨内侧平台的负重区以100-□m的间隔拾取三个切片，并用番红O-固绿色和HE染色。使用Image-Pro Plus 4定量每个载玻片中的骨赘面积和关节软骨厚度。TNFR 1-/-小鼠OA的手术诱导TNFR 1-/-及其野生型（WT）同窝仔通过内侧副韧带横断和内侧韧带切除术手术诱导发展OA。手术后4周，小鼠被安乐死。ResultsAt 52-60周龄，而骨赘的形成检测在胫骨平台的中前缘，无论基因型，骨赘面积显着增加TNFR 1-/-小鼠。此外，TNFR 1-/-小鼠胫骨平台内侧端区域的关节软骨厚度减少，表明TNFR 1缺乏促进了关节软骨细胞的骨化。接下来，我们通过诱导膝关节不稳定性来建立手术骨关节炎模型，比较了野生型和TNFR 1-/-小鼠的成年同窝仔之间的骨关节炎发展。组织学评价证实，TNFR 1缺乏导致骨赘形成显著加速。软骨破坏程度在TNFR 1-/-和WT之间几乎相当。新生小鼠骨骺软骨细胞的真实的时间PCR分析显示，与WT同窝小鼠相比，TNFR 1-/-小鼠中X型胶原（软骨细胞肥大的标志物）的mRNA表达显著升高。目前，我们正在挑选一些候选基因的目标TNFR 1的软骨细胞hypertrophy using cDNA microarray analysis.ConclusionEndogenous TNFR 1通过抑制X型胶原合成对骨赘形成有保护作用。少