Inhibition of RA synovial fibroblasts invasion by green tea polyphenols

描述(由申请人提供)：补充和替代医学方法的使用在类风湿性关节炎(RA)患者中越来越受欢迎。RA是一种慢性炎症性疾病，影响大约1.0%的美国人口，是一个重大的健康和社会经济挑战。在RA中，活化的滑膜成纤维细胞通过黏附分子和趋化因子协同作用的网络入侵关节软骨和骨，从而诱导炎症和基质降解酶的释放。在我们的初步研究中，我们发现富含多酚的绿茶水提物(绿茶多酚，GTP)可抑制白介素1(IL-1)诱导的人RA滑膜成纤维细胞CC[单核细胞趋化蛋白1(MCP-1)]的产生，并调节T细胞的正常表达和分泌(RANTES)和CXC[IL-8和生长调节癌基因-a(Gro-a)]趋化因子，(Ii)环氧合酶-2(COX-2)的表达，以及(Iii)核因子-kBp65(NF-kBp65)的核转位。此外，GTP可显著阻断体外培养的RA滑膜成纤维细胞由结构性和IL-1β诱导的基质金属蛋白酶-2(MMP2)活性。这项提议充分利用了这些新颖的观察结果。这项应用中提出的工作的中心假设是，GTP通过阻断趋化因子的产生、细胞黏附和基质金属蛋白酶的激活来抑制RA滑膜成纤维细胞的侵袭。在特定的目标1中，我们将利用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。在特定的目标2中，我们将测试GTP是否通过阻断IL-1诱导的(A)细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)等黏附分子的表达，(B)参与MMP-2、-3和-13激活的信号通路，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是否有效地抑制趋化因子介导的细胞迁移和RA滑膜成纤维细胞中基质金属蛋白酶的激活。这些研究结果将为GTP作为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.