Sumariwalla P F, Palmer C D, Pickford L B, Feldmann M, Foxwell B M J, Brennan F M
Kennedy Institute of Rheumatology, Imperial College London, London, UK.
Rheumatology (Oxford). 2009 Jan;48(1):32-8. doi: 10.1093/rheumatology/ken398. Epub 2008 Nov 16.
To evaluate the clinical efficacy of a novel synthetic peroxisome proliferator-activated receptor gamma (PPAR-gamma) agonist, CLX-090717, in several in vitro cell culture systems and murine CIA, an experimental model of RA.
Peripheral blood monocytes purified by elutriation, and rheumatoid synovial cells isolated from clinical tissue were cultured with CLX-090717 and TNF-alpha release was measured. Molecular mechanism of action was analysed by western blotting and electrophoretic mobility shift assay. Thioglycollate-elicited murine peritoneal macrophages were cultured with CLX-090717 and lipopolysaccharide (LPS)-induced TNF-alpha release was assayed. Therapeutic studies were done in mice with established arthritis by evaluating clinical parameters and histology. In addition, type II collagen response of lymphocytes from mice with CIA was examined.
CLX-090717 significantly inhibited spontaneous TNF-alpha release by RA synovial membrane cells, as well as LPS-induced TNF-alpha release from human and murine monocytic cells. Inhibition of TNF-alpha in monocytes was mediated partially through a nuclear factor-kappaB (NF-kappaB)-dependent pathway, as judged by sustained levels of IkappaBalpha in cytosolic extracts and a reduced level of LPS-induced NF-kappaB activity in nuclear extracts. CLX-090717 reduced clinical signs of arthritis and damage to joint architecture when administered therapeutically to arthritic mice. Mechanisms of action in CIA involved the reduction in proliferation of arthritic lymphocytes to antigen in vitro as well as reduced TNF-alpha release.
Our data suggest that the synthetic compound CLX-090717 has potential as a small molecular weight anti-inflammatory therapeutic for chronic inflammatory conditions.
在几种体外细胞培养系统和类风湿性关节炎（RA）的实验模型——小鼠胶原诱导性关节炎（CIA）中，评估新型合成过氧化物酶体增殖物激活受体γ（PPAR-γ）激动剂CLX-090717的临床疗效。
通过淘析法纯化外周血单核细胞，从临床组织中分离类风湿性滑膜细胞，用CLX-090717进行培养，并检测肿瘤坏死因子-α（TNF-α）的释放。通过蛋白质免疫印迹法和电泳迁移率变动分析来分析作用的分子机制。用CLX-090717培养巯基乙酸诱导的小鼠腹腔巨噬细胞，并检测脂多糖（LPS）诱导的TNF-α释放。通过评估临床参数和组织学，对已患关节炎的小鼠进行治疗研究。此外，还检测了CIA小鼠淋巴细胞对II型胶原的反应。
CLX-090717显著抑制RA滑膜细胞自发释放TNF-α，以及LPS诱导的人和小鼠单核细胞释放TNF-α。单核细胞中TNF-α的抑制部分是通过核因子-κB（NF-κB）依赖性途径介导的，这可通过细胞溶质提取物中IκBα的持续水平以及核提取物中LPS诱导的NF-κB活性降低来判断。当对患关节炎的小鼠进行治疗性给药时，CLX-090717可减轻关节炎的临床症状和关节结构损伤。CIA中的作用机制包括体外关节炎淋巴细胞对抗原增殖的减少以及TNF-α释放的减少。
我们的数据表明，合成化合物CLX-090717有潜力作为一种治疗慢性炎症性疾病的小分子量抗炎药物。
