Zaichko K, Stanislavchuk M, Zaichko N, Khomenko V
National Pirogov Memorial Medical University, Vinnytsya, Ukraine.
Georgian Med News. 2020 Feb(299):93-100.
Despite significant progress in treatment of rheumatoid arthritis (RA), a considerable part of patients remains resistant to the current therapy, apparently for the reasons of undefined mechanisms of its pathogenesis. Recently, the disturbances of circadian regulation of inflammatory processes in RA have been highlighted as important ones. Endothelial nitric oxide synthase (NOS3) and soluble toll-like receptors 2 (sTLR2) take part in the regulation of angiogenesis, osteoclastogenesis, immune responses but their circadian rhythms and predictive significance in RA patients are still unknown. Aim - to estimate the associations between efficacy of treatment and the circadian rhythms of NOS3 and sTLR2, and NOS3 polymorphism in females with rheumatoid arthritis, Ukraine. 97 RA patients (100% female) aged 46.3±8.89 years with disease duration 8.44±6.52 years were examined. All patients as a disease-modifying therapy received methotrexate (MTX) orally in a dose ≤15 mg/week, folic acid 5 mg/week, NSAIDs and corticosteroids (CS) ≤10 mg/day by prednisone. Doses of MTX, NSAIDs and CS were stable 4 weeks prior to the enrolment and during the whole period of study. The efficacy end points included DAS28, RAID and American College of Rheumatology response criteria (ACR20/50/70). Serum levels of NOS3 and sTLR2 were determined at 08:00 and 20:00 using Cloud-Clone Corp kits (USA). NOS3 T-786С polymorphism was determined by Real-Time PCR. The SPSS22 software package was used for statistical processing of the results. The study was performed in accordance to the bioethical standards. After 12-week treatment among RA patients were revealed 52.6% ACR 20 responders and 47.4% non-responders. Opposite diurnal variation of NOS3 and sTLR2 serum levels were found in RA patients. There were significant differences in NOS3/sTLR2 ratio at 08:00 accordingly to NOS3 T786C genotype. The disturbances in daily variability of NOS3 or sTLR2 serum levels were more significant in non-responders compare to responders. Decrease of NOS3/sTLR2 ratio was a predictor of non-response to treatment in RA patients (β=0.366, р=0.000). In RA patients the disturbances of circadian rhythms of endothelial nitric oxide synthase or toll-like receptors 2 expression are associated with an increase of resistance to disease-modifying therapy with methotrexate.
尽管类风湿关节炎（RA）的治疗取得了显著进展，但相当一部分患者对当前治疗仍有抵抗，显然是由于其发病机制不明。最近，RA中炎症过程昼夜调节的紊乱被认为是重要因素。内皮型一氧化氮合酶（NOS3）和可溶性Toll样受体2（sTLR2）参与血管生成、破骨细胞生成和免疫反应的调节，但其昼夜节律以及在RA患者中的预测意义仍不清楚。目的——评估乌克兰类风湿关节炎女性患者的治疗效果与NOS3和sTLR2昼夜节律以及NOS3基因多态性之间的关联。对97例年龄46.3±8.89岁、病程8.44±6.52年的RA患者（100%为女性）进行了检查。所有患者作为改善病情的治疗，口服甲氨蝶呤（MTX），剂量≤15mg/周，叶酸5mg/周，非甾体抗炎药（NSAIDs）和皮质类固醇（CS）按泼尼松计≤10mg/天。MTX、NSAIDs和CS的剂量在入组前4周及整个研究期间保持稳定。疗效终点包括疾病活动度评分28（DAS28）、类风湿关节炎疾病活动指数（RAID）和美国风湿病学会反应标准（ACR20/50/70）。使用美国Cloud-Clone Corp试剂盒在08:00和20:00测定血清中NOS3和sTLR2的水平。通过实时聚合酶链反应测定NOS3 T-786С基因多态性。结果采用SPSS22软件包进行统计处理。该研究按照生物伦理标准进行。经过12周治疗后，RA患者中显示52.6%达到ACR 20反应标准，47.4%未达到。在RA患者中发现NOS3和sTLR2血清水平存在相反的昼夜变化。根据NOS3 T786C基因型，08:00时NOS3/sTLR2比值存在显著差异。与反应者相比，未反应者中NOS3或sTLR2血清水平的每日变化紊乱更为显著。NOS3/sTLR2比值降低是RA患者治疗无反应的一个预测指标（β=0.366，p=0.000）。在RA患者中，内皮型一氧化氮合酶或Toll样受体2表达的昼夜节律紊乱与对甲氨蝶呤改善病情治疗的抵抗增加有关。
