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Development of Novel Therapeutic Strategies for Rheumatoid Arthritis utilizing the Extracellular Adenosine and Signaling via Cell Surface Adenosine Receptors.

利用细胞外腺苷和通过细胞表面腺苷受体的信号传导开发类风湿关节炎的新治疗策略。

基本信息

批准号：
11670447
负责人：
KOSHIBA Masahiro
金额：
$ 2.3万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

1) Effects of Extracellular Adenosine on Human Synovial Cells2-Chloroadenosine (2-CADO), an adenosine deaminase (ADA) resistant general Adenosine (Ado) agonist, induced the apoptosis on the fibroblast-like synoviocytes (FLS) from rheumatoid arthritis (RA) patients (RA-FLS) in a dose-dependent manner. Incubation of FLS with Ado resulted in the inhibition of proliferation as early as an hour, which was time- and concentration-dependent.2) Expression of Cell-surface Adenosine Receptors on FLSRT-PCR assay revealed that RA-FLS and two human FLS lines expressed all the four Adenosine Receptors (AdoR) mRNA (A_1, A_<2A>, A_<2B> and A_3). Functional analysis using the selective AdoR agonists and antagonists revealed that cAMP-increasing A_<2B> AdoR was functionally dominant on FLS.3) Molecular Mechanisms of Adenosine-mediated Apoptosis on FLSA potent ado transporter inhibitor NBT, but the selective AdoR antagonists, suppressed the RA-FLS apoptosis bar 2-CADO.Incubation with cell-permeable cAMP analog or co-incubation of Ado with ADA inhibitor also failed to inhibit the apoptosis, while general caspase inhibitor z-VAD-fmk was capable of blocking the apoptosis. These data indicated the machinery that the exogenous 2-CADO after entering into the cells activated the caspases and induced the apoptotic cell death.4) Anti-rheumatic Effect of Adenosine Transporter Inhibitors in vivoThe results mentioned above opened the possibility of the ado transporter inhibitors as novel ant-rheumatic drugs. We have started to test this in rat adjuvant arthritis model. The preliminary results were that the systemic administration of NBT appears similarly or even more effective than methotrexate. Thus the further in vivo investigation of the anti-rheumatic effects of NBT and other nucleoside transporters will open a new therapeutic strategy of RA.

1)细胞外腺苷对人滑膜细胞的作用腺苷脱氨酶（ADA）拮抗剂2-氯腺苷（2-CADO）以剂量依赖方式诱导类风湿关节炎（RA）成纤维样滑膜细胞（RA-FLS）凋亡。（2）细胞表面腺苷受体表达的FLSRT-PCR检测结果显示，RA-FLS和两株人FLS细胞均表达腺苷受体（A_1、<2A>A_3<2B>、A_1）mRNA。3<2B>）腺苷介导的细胞凋亡的分子机制：FLSA强的ado转运体抑制剂NBT抑制RA-FLS的凋亡，而选择性的ADDR拮抗剂抑制RA-FLS的凋亡，但2-CADO抑制作用不明显，细胞可渗透的cAMP类似物孵育或Ado与ADA抑制剂共孵育也不能抑制凋亡，而一般的caspase抑制剂z-VAD-fcadine则能阻断凋亡。这些数据表明外源性2-CADO进入细胞后激活caspase并诱导细胞凋亡的机制。4）腺苷转运体抑制剂的体内抗风湿作用上述结果为腺苷转运体抑制剂作为新型抗风湿药物提供了可能性。我们已经开始在大鼠佐剂性关节炎模型中测试这一点。初步结果是NBT的全身给药似乎与甲氨蝶呤相似或甚至更有效。因此，进一步研究NBT和其他核苷转运体的抗风湿作用将为RA的治疗开辟新的途径。