Inhibition of RA synovial fibroblasts invasion by green tea polyphenols

描述（由申请人提供）：使用补充和替代医学方法在类风湿性关节炎（RA）患者中越来越受欢迎。类风湿性关节炎是一种慢性炎症性疾病，影响约1.0%的美国人口，是一个重大的健康和社会经济挑战。在类风湿性关节炎中，激活的滑膜成纤维细胞通过粘附分子和趋化因子的网络侵入关节软骨和骨骼，共同诱导炎症和基质降解酶的释放。在我们的初步研究中，我们发现绿茶(Camellia sinensis；绿茶多酚（GTP）阻断白细胞介素-1 （IL-1）诱导的(i) CC[单核细胞趋化蛋白1 （MCP-1）的产生，并调节正常T细胞表达和分泌（RANTES）和CXC [IL-8和生长调节癌基因-a (Gro-a)]趋化因子的激活，（ii）环氧化酶-2 （COX-2）的表达，以及（iii）体外人RA滑膜成纤维细胞核因子- kbp65 （NF-kBp65）的核易位。此外，GTP治疗可显著阻断体外RA滑膜成纤维细胞中组成型和IL-1诱导的基质金属蛋白酶-2 （MMP-2）活性。这个建议利用了这些新颖的观察结果。本研究的中心假设是GTP会通过阻断趋化因子的产生、细胞粘附和MMP的激活来抑制RA滑膜成纤维细胞的侵袭。在特定的目的1中，我们将使用RA滑膜成纤维细胞，研究GTP是否通过抑制/改变IL-1诱导的信号通路和趋化因子受体的表达来抑制趋化因子的产生。在特定目的2中，我们将测试GTP是否通过阻断IL-1诱导的(a)粘附分子如细胞间粘附分子-1 （ICAM-1）和血管细胞粘附分子-1 （VCAM-1）的表达，(b)与MMP-2、-3和-13激活相关的信号通路，以及(c) COX-2的表达来抑制RA滑膜成纤维细胞的侵袭。在具体目标3中，我们将研究GTP是否有效抑制趋化因子介导的RA滑膜成纤维细胞的细胞迁移和MMP激活。这些研究结果将为GTP作为治疗实体靶向趋化因子及其受体介导的滑膜侵袭在啮齿动物关节炎模型和人类RA中的体内研究奠定基础。

DESCRIPTION (provided by applicant): The use of complementary and alternative medicine approaches is becoming increasingly popular with rheumatoid arthritis (RA) patients. RA is a chronic inflammatory disease affecting approximately 1.0% of the American population and is a significant health and socio-economic challenge. In RA, activated synovial fibroblasts invade the articular cartilage and bone by a network of adhesion molecules and chemokines working in concert to induce inflammation and the release of matrix-degrading enzymes. In our preliminary studies, we found that the polyphenol-rich aqueous fraction of green tea (Camellia sinensis; green tea polyphenols, GTP) blocked interleukin-1¿ (IL-1¿)-induced (i) production of the CC [monocyte chemotactic protein 1 (MCP-1) and regulated upon activation normally T cells expressed and secreted (RANTES] and CXC [IL-8 and growth regulated oncogene-a (Gro-a)] chemokines, (ii) cyclooxygenase-2 (COX-2) expression, and (iii) nuclear translocation of nuclear factor-kBp65 (NF-kBp65) in human RA synovial fibroblasts in vitro. In addition, treatment with GTP significantly blocked both constitutive- and IL-1¿-induced matrix metalloproteinase-2 (MMP-2) activity in RA synovial fibroblasts in vitro. This proposal capitalizes on these novel observations. The central hypothesis of the work proposed in this application is that GTP will inhibit RA synovial fibroblast invasion by blocking chemokine production, cell adhesion, and MMP activation. In specific aim 1, using RA synovial fibroblasts, we will study whether GTP inhibits chemokine production by inhibiting/altering IL-1¿ induced signaling pathways and the expression of chemokine receptors. In specific aim 2, we will test whether GTP suppresses RA synovial fibroblast invasion by blocking IL-1¿ induced (a) expression of adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), (b) signaling pathways implicated in MMP-2, -3, and -13 activation, and (c) COX-2 expression. In specific aim 3, we will study whether GTP is effective in inhibiting chemokine mediated cell migration and MMP activation in RA synovial fibroblasts. Results from these studies will lay the foundation for in vivo studies for GTP as therapeutic entity to target chemokines and their receptor mediated synovial invasion in rodent arthritis models and in human RA.