Functional analysis of plasminogen related genes in cartilage metabolism

本研究的目的是阐明PRGB和PRPB在类风湿关节炎等关节炎中的作用。我们研究了PRGB的表达和PRPB的主要组成部分，滑膜和软骨，在类风湿关节炎密切涉及血管生成的作用。PRGB在体外正常条件下在滑膜细胞中不表达，并且表达不被诱导，即使当细胞用细胞因子（IL-1和bFGF）刺激以模拟类似于类风湿性关节炎的条件时。在软骨中，PRGB在正常条件下在软骨细胞中不表达，但通过细胞因子刺激增强表达。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，但当受到刺激时，例如在炎症期间，表达增强，尽管机制仍不清楚。为了研究PRGB的作用，我们制备了重组P 关于我们 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。当用细胞因子（IL-1和bFGF）刺激时，滑膜细胞增殖。但PRPB对细胞增殖无抑制作用。因此，我们集中在假设VEGF可以强烈诱导新血管形成的炎症滑膜在RA，并研究PRPB对VEGF表达的影响。首先，我们将PRPB加入到细胞因子（IL-1和bFGF）刺激的滑膜细胞和软骨细胞中，从基因和蛋白水平研究VEGF表达的变化。PRPB在基因和蛋白水平上显著抑制滑膜细胞中的VEGF表达，并在基因水平上抑制软骨中的VEGF表达。对于体内研究，使用假定代表类风湿性关节炎病理的CIA小鼠。与用PBS处理的对照组相比，PRPB处理组的爪厚度随时间显著降低。此外，microCT显示骨侵蚀和破坏减少。组织学上，滑膜生长和骨破坏均减少，并且用抗CD 31抗体免疫染色关节滑膜组织中的血管内皮细胞时，CD 31阳性血管内皮细胞的数量也减少。提示PRPB治疗组关节炎得到抑制，血管生成受到抑制。关于PRPB对血管内皮细胞的作用，通过整合素发挥抑制作用。还表明抑制滑膜组织产生的VEGF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，仍有待研究。少

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