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Functional analysis of plasminogen related genes in cartilage metabolism

软骨代谢中纤溶酶原相关基因的功能分析

基本信息

批准号：
18591677
负责人：
MORIOKA Hideo
金额：
$ 2.23万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591677/
关键词：
Joint disease Rheumatoid arthritis Osteoarthritis Bone and Cartilage metabolism Angiogenesis Cartilage Synovium Synovitis

项目摘要

The purpose of this study is to clarify the role of PRGB and PRPB in arthritis joint such as RA. We investigated the expression of PRGB and the role of PRPB in the main components, synovium and cartilage, in rheumatoid arthritis closely involving angiogenesis. PRGB was not expressed in synoviocytes under normal conditions in vitro, and expression was not induced, even when the cells were stimulated with cytokines (IL-1 and bFGF) to mimic a condition similar to rheumatoid arthritis. In cartilage, PRGB was not expressed in chondrocytes under normal conditions, but the expression was enhanced by cytokine stimulation. On ISH, PRGB was not expressed in the synovial membrane, but was partially expressed in joint cartilage with RA. These findings suggested that PRGB is not expressed in normal joint cartilage, but that the expression is enhanced when stimulated, such as during inflammation, although the mechanism remains unclear. To investigate the action of PRGB, we prepared the recombinant P … More RGB protein (PRPB). In RA, the influence of PRPB on synoviocytes was investigated because synovium growth is involved in the pathogenesis. Synoviocytes proliferated when stimulated with cytokines (IL-1 and bFGF). However, PRPB did not inhibit the proliferation. Thus, we focused on the hypothesis that VEGF could strongly induce new blood vessel formation in inflammatory synovium in RA, and investigated the influence of PRPB on VEGF expression. Firstly, we added PRPB to synoviocytes and chondrocytes stimulated with cytokines (IL-1 and bFGF), and investigated the changes in VEGF expression at the gene and protein levels. PRPB significantly inhibited VEGF expression in synoviocytes at both the gene and protein levels, and in cartilage at the gene level. For in vivo study, CIA mice that are presumed to represent the pathology of rheumatoid arthritis were used. The paw thickness was significantly reduced over the time in the PRPB treatment group, compared to that in the control group treated with PBS. In addition, bone erosion and destruction were reduced on micro CT. Histopathologically, synovium growth and bone destruction were both reduced, and the number of CD31-positive vascular endothelial cells was also decreased on immunostaining of vascular endothelial cells in the articular synovial tissue with anti-CD31 antibody. It was suggested that arthritis was inhibited and angiogenesis was suppressed in the PRPB treatment group. Regarding the action of PRPB on vascular endothelial cells, the inhibitory effect was exerted via integrin. It was also suggested that inhibition of VEGF produced by the synovial tissue suppressed angiogenesis. The mechanism of action of the inhibition of VEGF by PRPB remains to be investigated. This study confirmed that PRPB inhibits arthritis by suppressing angiogenesis, but many of the details of the mechanism of action are unclear and remain to be investigated. Less

本研究的目的是阐明PRGB和PRPB在RA等关节炎关节中的作用。我们研究了与血管生成密切相关的类风湿性关节炎的主要成分滑膜和软骨中 PRGB 的表达以及 PRPB 的作用。在体外正常条件下，PRGB 在滑膜细胞中不表达，即使用细胞因子（IL-1 和 bFGF）刺激细胞以模拟类似于类风湿性关节炎的情况，也不会诱导表达。在软骨中，PRGB在正常情况下在软骨细胞中不表达，但在细胞因子刺激下表达增强。在 ISH 上，PRGB 在滑膜中不表达，但在 RA 的关节软骨中部分表达。这些发现表明，PRGB 在正常关节软骨中不表达，但在受到刺激时（例如在炎症期间）表达会增强，尽管其机制尚不清楚。为了研究 PRGB 的作用，我们制备了重组 P RGB 蛋白（PRPB）。在 RA 中，研究了 PRPB 对滑膜细胞的影响，因为滑膜生长参与了发病机制。当细胞因子（IL-1 和 bFGF）刺激时，滑膜细胞增殖。然而，PRPB并不抑制增殖。因此，我们关注VEGF可以强烈诱导RA炎症滑膜中新血管形成的假设，并研究PRPB对VEGF表达的影响。首先，我们将PRPB添加到细胞因子（IL-1和bFGF）刺激的滑膜细胞和软骨细胞中，并在基因和蛋白水平上研究VEGF表达的变化。 PRPB 在基因和蛋白质水平上显着抑制滑膜细胞中的 VEGF 表达，并在基因水平上显着抑制软骨中的 VEGF 表达。对于体内研究，使用了被认为代表类风湿性关节炎病理的 CIA 小鼠。与用 PBS 治疗的对照组相比，PRPB 治疗组的爪子厚度随着时间的推移显着减小。此外，显微CT显示骨侵蚀和破坏也减少了。组织病理学上，用抗CD31抗体对关节滑膜组织中的血管内皮细胞进行免疫染色，滑膜生长和骨破坏均减少，并且CD31阳性血管内皮细胞的数量也减少。表明PRPB治疗组关节炎被抑制并且血管生成被抑制。 PRPB对血管内皮细胞的作用，是通过整合素发挥抑制作用的。还表明抑制滑膜组织产生的 VEGF 可以抑制血管生成。 PRPB抑制VEGF的作用机制仍有待研究。这项研究证实PRPB通过抑制血管生成来抑制关节炎，但作用机制的许多细节尚不清楚，有待研究。较少的