Varani Katia, De Mattei Monica, Vincenzi Fabrizio, Tosi Alice, Targa Martina, Masieri Federica F, Pellati Agnese, Massari Leo, Borea Pier Andrea
Department of Clinical and Experimental Medicine, University of Ferrara, 44100 Ferrara, Italy.
Cell Physiol Biochem. 2010;25(2-3):325-36. doi: 10.1159/000276565. Epub 2010 Jan 12.
BACKGROUND/AIMS: P2X receptors are membrane ion channels activated by extracellular adenosine 5'-triphosphate (ATP) which contribute to various physiological processes. The present study describes in synovial fibroblasts (SFs) obtained from osteoarthritis (OA) patients and in SW 982 cells derived from human synovial sarcoma a pharmacological characterization of P2X(1) and P2X(3) receptors implicated in the modulation of inflammatory processes in joint diseases.
mRNA, western blotting, saturation and competition binding experiments were used to characterize purinergic receptors. From a functional point of view nuclear factor kappaB (NF-kappaB) activation, tumour necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6) and prostaglandin E(2) (PGE(2)) production were evaluated by means of enzyme-linked immunosorbent assays.
P2X(1) and P2X(3) receptors were present with high affinity and density. Selected purinergic agonists and antagonists exhibited a different thermodynamic behavior. P2X(1) receptors showed an anti-inflammatory effect reducing NF-kappaB activation and TNF-alpha release whilst P2X(3) receptors mediated opposite response. No effect was mediated by P2X(1) and P2X(3) receptors on IL-6 and PGE(2) production.
SFs from OA patients and SW 982 cells similarly express P2X(1) and P2X(3) receptors which are able to modulate in opposite way some functional responses closely associated with inflammation suggesting that purinergic receptors may represent a potential target in therapeutic anti-inflammatory joint interventions.
背景/目的：P2X受体是由细胞外三磷酸腺苷（ATP）激活的膜离子通道，参与多种生理过程。本研究描述了从骨关节炎（OA）患者获得的滑膜成纤维细胞（SFs）以及源自人滑膜肉瘤的SW 982细胞中，与关节疾病炎症过程调节相关的P2X(1)和P2X(3)受体的药理学特性。
采用mRNA、蛋白质印迹、饱和与竞争结合实验来表征嘌呤能受体。从功能角度，通过酶联免疫吸附测定法评估核因子κB（NF-κB）激活、肿瘤坏死因子-α（TNF-α）、白细胞介素6（IL-6）和前列腺素E(2)（PGE(2)）的产生。
P2X(1)和P2X(3)受体以高亲和力和密度存在。选定的嘌呤能激动剂和拮抗剂表现出不同的热力学行为。P2X(1)受体显示出抗炎作用，可降低NF-κB激活和TNF-α释放，而P2X(3)受体介导相反的反应。P2X(1)和P2X(3)受体对IL-6和PGE(2)的产生无影响。
OA患者的SFs和SW 982细胞同样表达P2X(1)和P2X(3)受体，它们能够以相反的方式调节一些与炎症密切相关的功能反应，这表明嘌呤能受体可能是治疗性抗炎关节干预的潜在靶点。
