Investigation of the mechanism of joint destruction in inflammatory process and the development of its prophyractic treatement by cellular and molecular biological approach

为探讨粘附分子（VLA-4、VLA-5）、基质金属蛋白酶（MMP）及基质金属蛋白酶组织抑制剂（TIMP）在类风湿关节炎（RA）滑膜中的定位，采用免疫组织化学方法对RA软骨-脂膜连接进行了研究。VLA-5在与细胞外软骨基质接触的滑膜细胞的细胞表面上强烈表达，这表明VLA-5和构成基质的纤连蛋白的相互作用可能在类风湿性滑膜中血管翳的形成和侵袭中是重要的。软骨-脂膜连接处MMP-1和MMP-3均呈阳性表达，与VLA-5抗原接近，而TIMP-1表达较弱。本研究还观察了炎症刺激对滑膜成纤维细胞粘附分子表达的影响，以探讨抑制粘附分子表达及其功能，从而抑制RA关节破坏的可能性。炎性细胞因子如白细胞介素-1 β增强了人巨噬细胞系U-937与RA滑膜成纤维细胞的细胞附着。虽然粘附分子如ICAM-1和VCAM-1的表达在炎性细胞因子的刺激下增强，但VLA-5的表达一直稳定。这些结果表明，VLA-5没有受到炎症刺激和VLA-5的直接抗体的功能抑制的定量影响可能是必要的RA治疗的调节粘附分子的表达和细胞附着。

In order to examine the localization of adhesion molecule (VLA-4, VLA-5), matrix metalloproteinase (MMP) and tissue inhibitor of matrix metalloproteinase (TIMP) in rheumatoid synovium, the cartilage-pannusjunction in rheumatoid arthritis (RA) were explored immunohistochemically. VLA-5 is strongly expressed on the cell surface of synovial cells contacting extracellular cartilage matrix, which is suggesting that interaction of VLA-5 and fibronectin constituting the matrix might be important in pannus formation and invasion in rheumatoid synovium. Both MMP-1 and MMP-3 are positively stained closed to the VLA-5 antigen in cartilage-pannusjunction, whereas the expression of TIMP-1 was very weak. These results may indicate that imbalance of MMP and TIMP in cartilage-pannus junction may accelerate the invasion of pannus on articular cartilage lead to joint destruction.Modulation of adhesion molecule expression on cultured synovial fibroblasts by inflammatory stimulation were also examined in order to elucidate the possibilities of the suppression of joint destruction in RA by inhibiting the expression of adhesion molecule and its function. Inflammatory cytokines such as interleukin-1β has augmented the cell attachment of human macrophage cell line U-937 to RA synovial fibroblasts. While the expression of adhesion molecules such as 1CAM-1 and VCAM-1 were enhanced by stimulation of inflammatory cytokines, the expression of VLA-5 has been stable. These results suggest that VLA-5 has not been influenced quantitively by inflammatory stimulation and functional inhibition of VLA-5 directly by antibodies might be necessary for the RA treatment in terms of the modulation of adhesion molecule expression and cell attachment.