Shibata-Nozaki Toshiko, Ito Hiroshi, Mitomi Hirofumi, Akaogi Jun, Komagata Tatsuya, Kanaji Toshiya, Maruyama Takayuki, Mori Toshihito, Nomoto So, Ozaki Shoichi, Yamada Hidehiro
Department of Internal Medicine, St. Marianna University School of Medicine, Kawasaki, Japan.
Arthritis Rheum. 2011 Sep;63(9):2595-605. doi: 10.1002/art.30428.
We recently developed an ex vivo cellular model of pannus, the aberrant overgrowth of human synovial tissue. This study was undertaken to use that model to investigate the role of prostaglandin E2 (PGE2) and its receptor subtypes in the development of pannus growth and osteoclast activity in rheumatoid arthritis (RA).
Inflammatory cells that infiltrated pannus from patients with RA were collected without enzyme digestion and designated synovial tissue-derived inflammatory cells. Their single-cell suspensions were cultured in medium alone to observe an aberrant overgrowth of inflammatory tissue in vitro. Levels of cytokines produced in culture supernatants were measured using enzyme-linked immunosorbent assay kits. Osteoclast activity was assessed by the development of resorption pits in calcium phosphate-coated slides.
Primary culture of the synovial tissue-derived inflammatory cells resulted in spontaneous reconstruction of inflammatory tissue in vitro within 4 weeks, during which tumor necrosis factor α, PGE2, macrophage colony-stimulating factor, and matrix metalloproteinase 9 were produced in the supernatant. This aberrant overgrowth was inhibited by antirheumatic drugs including methotrexate and infliximab. On calcium phosphate-coated slides, synovial tissue-derived inflammatory cells showed numerous resorption pits. In the presence of inhibitors of endogenous prostanoid production such as indomethacin and NS398, exogenous PGE1 and EP4-specific agonists significantly inhibited all these activities of synovial tissue-derived inflammatory cells in a dose-dependent manner. Addition of indomethacin, NS398, or EP4-specific antagonist resulted in the enhancement of these cells' activities. EP2-specific agonist had a partial effect, while EP1- and EP3-specific agonists had no significant effects.
These results suggest that endogenous PGE2 produced in rheumatoid synovium negatively regulates aberrant synovial overgrowth and the development of osteoclast activity via EP4.
我们最近建立了一种体外细胞模型——血管翳，即人类滑膜组织的异常过度生长。本研究旨在利用该模型研究前列腺素E2（PGE2）及其受体亚型在类风湿关节炎（RA）血管翳生长和破骨细胞活性发展中的作用。
从类风湿关节炎患者的血管翳中收集浸润的炎性细胞，无需酶消化，命名为滑膜组织来源的炎性细胞。将其单细胞悬液单独培养于培养基中，以观察体外炎性组织的异常过度生长。使用酶联免疫吸附测定试剂盒测量培养上清液中产生的细胞因子水平。通过磷酸钙包被载玻片上吸收陷窝的形成来评估破骨细胞活性。
滑膜组织来源的炎性细胞原代培养在4周内导致体外炎性组织的自发重建，在此期间上清液中产生肿瘤坏死因子α、PGE2、巨噬细胞集落刺激因子和基质金属蛋白酶9。这种异常过度生长受到包括甲氨蝶呤和英夫利昔单抗在内的抗风湿药物的抑制。在磷酸钙包被的载玻片上，滑膜组织来源的炎性细胞显示出大量吸收陷窝。在内源性前列腺素生成抑制剂如吲哚美辛和NS398存在的情况下，外源性PGE1和EP4特异性激动剂以剂量依赖性方式显著抑制滑膜组织来源的炎性细胞的所有这些活性。添加吲哚美辛、NS398或EP4特异性拮抗剂导致这些细胞活性增强。EP2特异性激动剂有部分作用，而EP1和EP3特异性激动剂无显著作用。
这些结果表明，类风湿滑膜中产生的内源性PGE2通过EP4负向调节滑膜异常过度生长和破骨细胞活性的发展。
