Loredo G A, Benton H P
University of California, Davis 95616, USA.
Arthritis Rheum. 1998 Feb;41(2):246-55. doi: 10.1002/1529-0131(199802)41:2<246::AID-ART8>3.0.CO;2-I.
To pharmacologically and functionally characterize calcium-mobilizing purine receptors on adherent human rheumatoid synovial cells.
Fura-2-loaded synovial cells were screened for changes in cytosolic calcium concentration after the addition of purine receptor agonists. Release of interleukin-1 (IL-1) and prostaglandin E2 (PGE2) was assessed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay, respectively. The effect of IL-1 prestimulation on purine-mediated PGE2 release was determined.
ATP (1-100 microM) and UTP (1-100 microM), but not 2-methylthio-ATP or adenosine, stimulated mobilization of calcium from intracellular stores in synovial cells. ATP and UTP stimulated a small, but significant, increase in PG release from resting synoviocytes and a dramatic increase in PG release from synoviocytes prestimulated with recombinant human IL-1alpha. Neither ATP nor UTP stimulated synoviocyte release of IL-1 as measured by specific ELISA. The effects of ATP and UTP on PG secretion were mimicked by phorbol 12-myristate 13-acetate and thapsigargin, and blocked by BAPTA buffering of cytosolic calcium.
Adherent human rheumatoid synovial cells mobilize intracellular calcium via a P2U-like purine receptor. P2U receptor agonists stimulate PGE2 release from synoviocytes, an effect that is greatly enhanced by IL-1alpha prestimulation and blocked by intracellular calcium buffering.
从药理学和功能方面对人类风湿性滑膜贴壁细胞上的钙动员嘌呤受体进行特性分析。
在添加嘌呤受体激动剂后，对负载Fura-2的滑膜细胞进行胞质钙浓度变化的筛选。分别通过酶联免疫吸附测定（ELISA）和放射免疫测定评估白细胞介素-1（IL-1）和前列腺素E2（PGE2）的释放。确定IL-1预刺激对嘌呤介导的PGE2释放的影响。
ATP（1 - 100微摩尔）和UTP（1 - 100微摩尔），而非2-甲硫基-ATP或腺苷，刺激滑膜细胞内钙库释放钙。ATP和UTP刺激静息滑膜细胞中PG释放少量但显著增加，而在经重组人IL-1α预刺激的滑膜细胞中PG释放显著增加。通过特异性ELISA测定，ATP和UTP均未刺激滑膜细胞释放IL-1。佛波醇肉豆蔻酸酯12-乙酸酯和毒胡萝卜素模拟了ATP和UTP对PG分泌的影响，并被胞质钙的BAPTA缓冲所阻断。
人类风湿性滑膜贴壁细胞通过一种P2U样嘌呤受体动员细胞内钙。P2U受体激动剂刺激滑膜细胞释放PGE-2，IL-1α预刺激可极大增强该效应，而细胞内钙缓冲可阻断该效应。
