Division of Rheumatology and Allergology, Department of Internal Medicine, St. Marianna University School of Medicine, 2-16-1 Sugao, Miyamae-ku, Kawasaki, 216-8511, Japan,
Mod Rheumatol. 2013 Nov;23(6):1069-75. doi: 10.1007/s10165-012-0794-7. Epub 2012 Nov 25.
During isometric exercise, the synovial joint tissue is prone to hypoxia, which is further enhanced in the presence of synovial inflammation. Hypoxia is also known to induce inflammatory cascades, suggesting that periodic hypoxia perpetuates synovitis in rheumatoid arthritis. We previously established an ex vivo cellular model of rheumatoid arthritis using the synovial tissue-derived inflammatory cells, which reproduced aberrant synovial overgrowth and pannus-like tissue development in vitro. Using this model, we investigated the regulatory mechanism of synovial cells against hypoxia in rheumatoid arthritis.
Inflammatory cells that infiltrated synovial tissue from patients with rheumatoid arthritis were collected without enzyme digestion, and designated as synovial tissue-derived inflammatory cells. Under normoxia or periodic hypoxia twice a week, their single-cell suspension was cultured in medium alone to observe an aberrant overgrowth of inflammatory tissue in vitro. Cytokines produced in the culture supernatants were measured by enzyme-linked immunosorbent assay kits.
Primary culture of the synovial tissue-derived inflammatory cells under periodic hypoxia resulted in the attenuation of the spontaneous growth of inflammatory tissue in vitro compared to the culture under normoxia. Endogenous prostaglandin E2 (PGE2) production was enhanced under periodic hypoxia. When endogenous PGE2 was blocked by indomethacin, the aberrant tissue overgrowth was more enhanced under hypoxia than normoxia. Indomethacin also enhanced the production of tumor necrosis factor-α (TNF-α), macrophage colony-stimulating factor (M-CSF), and matrix metalloproteinase-9 (MMP-9) under periodic hypoxia compared to normoxia. The EP4-specific antagonist reproduced the effect of indomethacin. Exogenous PGE1 and EP4-specific agonist effectively inhibited the aberrant overgrowth and the production of the inflammatory mediators under periodic hypoxia as well as normoxia.
The enhancing effect of periodic hypoxia on the aberrant overgrowth of rheumatoid synovial tissue was effectively down-regulated by the simultaneously induced endogenous PGE2.
在等长运动过程中,滑液关节组织容易缺氧,而在存在滑膜炎症的情况下,这种缺氧情况会进一步加剧。缺氧也已知会引发炎症级联反应,这表明周期性缺氧会使类风湿关节炎中的滑膜炎持续存在。我们之前使用滑膜组织来源的炎症细胞建立了类风湿关节炎的体外细胞模型,该模型在体外再现了异常的滑膜过度生长和类似肉芽组织的组织发育。使用该模型,我们研究了类风湿关节炎中滑膜细胞对缺氧的调节机制。
从类风湿关节炎患者的滑膜组织中收集未经过酶消化的炎症细胞,并将其命名为滑膜组织来源的炎症细胞。在常氧或每周两次周期性缺氧条件下,将其单细胞悬液在培养基中单独培养,以观察体外炎症组织的异常过度生长。通过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。
周期性缺氧下的滑膜组织来源的炎症细胞原代培养导致体外炎症组织的自发生长相对于常氧条件下减弱。内源性前列腺素 E2(PGE2)的产生在周期性缺氧下增强。当用吲哚美辛阻断内源性 PGE2 时,缺氧条件下的异常组织过度生长比常氧条件下更明显。与常氧相比,吲哚美辛还增强了周期性缺氧下肿瘤坏死因子-α(TNF-α)、巨噬细胞集落刺激因子(M-CSF)和基质金属蛋白酶-9(MMP-9)的产生。EP4 特异性拮抗剂再现了吲哚美辛的作用。外源性 PGE1 和 EP4 特异性激动剂可有效抑制周期性缺氧和常氧条件下异常过度生长和炎症介质的产生。
周期性缺氧对类风湿关节炎滑膜组织异常过度生长的增强作用可通过同时诱导的内源性 PGE2 有效下调。