Angiogenic action of gliostatin in rheumatoid arthritis and its molecular mechanism

胶质抑素对类风湿性关节炎的血管生成作用及其分子机制

基本信息

批准号：
16591504
负责人：
NAGAYA Yuko
金额：
$ 1.73万
依托单位：
NAGOYA CITY UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591504/
关键词：
rheumatoid arthritis synoviocytes angiogenesis gliostatin VEGF DNA microarray system gene transfection

项目摘要

Neovascularization, proliferation of synovial cells, and mononuclear cell influx and activation are characteristic events observed in synovial joints in the pathohistology of rheumatoid arthritis (RA). We have previously measured the concentration of the angiogenic factor, gliostatin (GLS), in sera and synovial fluids of RA patients, and demonstrated for the first time an enormously high concentration in RA synovial fluids as well as in RA sera. Furthermore our previous study demonstrated that GLS acts as a cytokine to augment its own synthesis in fibroblast-like synoviocytes (FLSs) obtained from patients with RA through an autocrine mechanism. It should be noted that GLS additionally caused induction and extracellular secretion of matrix metalloproteinase (MMP)-1 and MMP-3 triggering cartilage degeneration. Recently we reported that intraarticular injection of rHuGLS to rabbit knees induced RA-like synovitis.The purpose of this study was to elucidate whether GLS/TP is involved in the regulation of the angiogenic cytokine vascular endothelial growth factor (VEGF) in rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLSs) from patients with RA were cultured and stimulated with recombinant human GLS (rHuGLS) and interleukin (IL)-1β. Immunohistochemistry showed that GLS/TP and VEGF were detectable in the synovial lining cells. In cultured FLSs, both VEGF mRNA and protein levels were markedly increased by rHuIL-1βtreatment. rHuGLS increased VEGF mRNA expression in a dose-dependent manner. We detected high concentrations of VEGF165 protein in culture supernatants from FLSs treated with rHuGLS (300ng/ml), which were comparable to GLS levels found in synovial fluid of RA patients. These findings indicate that GLS/TP and VEGF have synergistic effects on angiogenesis in rheumatoid synovitis, and that GLS/TP has a role in regulating VEGF.

新血管形成，滑膜细胞的增殖以及单核细胞的影响和激活是类风湿关节炎病原体（RA）中滑膜关节中观察到的特征事件。我们先前已经在RA患者的血清和滑液中测量了血管生成因子（GLS）的浓度，并首次证明了RA滑液以及RA Sera中极高的浓度。此外，我们先前的研究表明，GL是一种细胞因子，可以增强其在通过自分泌机制中从RA患者获得的成纤维细胞样的滑膜细胞（FLS）中的合成。应该注意的是，GL还引起基质金属蛋白酶（MMP）-1和MMP-3触发软骨变性的诱导和提取的细胞分泌。最近，我们报道说，关节内向兔膝盖对兔膝盖诱导的RA样滑膜炎。该研究的目的是阐明类风湿性关节炎（RA）中GLS/TP是否参与了血管生成细胞因子血管内皮生长因子（VEGF）的调节。培养RA患者的成纤维细胞样滑膜炎（FLS），并用重组人GL（Rhugls）和白介素（IL）-1β刺激。免疫组织化学表明，在滑膜细胞中可以检测到GLS/TP和VEGF。在培养的FLS中，rhuil-1β处理显着提高了VEGF mRNA和蛋白质水平。 rhugls以剂量依赖性方式增加了VEGF mRNA的表达。我们检测到来自用Rhugls（300ng/ml）处理的FLS的培养上清液中的高浓度VEGF165蛋白，这些蛋白与RA患者滑液中的GLS水平相当。这些发现表明，GLS/TP和VEGF对类风湿滑膜炎的血管生成具有协同作用，并且GLS/TP在调节VEGF中起作用。