Novel effects mediated by bradykinin and pharmacological characterization of bradykinin B2 receptor antagonism in human synovial fibroblasts
Novel effects mediated by bradykinin and pharmacological characterization of bradykinin B2 receptor antagonism in human synovial fibroblasts
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DOI：
10.1111/j.1476-5381.2009.00511.x
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发表时间：
2009-12-01
影响因子：
7.3
通讯作者：
Maggi, C. A.
中科院分区：
文献类型：
--
作者：
Bellucci, F.;Cucchi, P.;Maggi, C. A.
背景与目的：缓激肽（BK）和B-2受体参与骨关节炎（OA）的病理生理过程，滑膜炎是OA的标志之一。在这里，选择性B-2受体拮抗剂MEN 16132和艾替班特已在人滑膜cells.Experimental approach：放射性配体和功能研究（磷酸肌醇（IP）的积累，白细胞介素（IL）-6和IL-8的释放）在培养的滑膜cells.Key results：[3 H]-BK饱和度研究表明受体密度（B-max）和Kd值分别为121 550个位点/细胞和1.14 nM。在滑膜细胞中，MEN 16132（pK（i）8.9）的效力是艾替班特（pK（i）8.4）的三倍。两种拮抗剂在BK诱导的IP试验中均显示出竞争性拮抗作用（对照EC 50 0.45 nM），pK（B）值为9.9（MEN 16132）和8.1（艾替班特）。与BK孵育24小时诱导IL-6（EC 50 216 nM）和IL-8（EC 50 53 nM）释放。MEN 16132（IL-6：pIC（50）8.1; IL-8：pIC（50）8.4）和艾替班特（IL-6：pIC（50）6.6; IL-8：pIC（50）6.7）完全阻止了这种BK诱导的释放。吲哚美辛不影响BK诱导的基础或IL-6/IL-8释放，而去甲二氢愈创木酸降低基础释放，尽管BK仍然增加IL-6和IL-8的产生。BK诱导的IL-8释放减弱的抑制剂磷脂酶C（U 73122），p38（SB 203580），JNK（SP 600125），ERK 1/2（PD 98059）MAPK，磷酸肌醇3-激酶（LY 294002），NF-κ B（BAY-117085）和糖皮质激素dexamethason.Conclusions和影响：缓激肽通过B-2受体可以参与滑膜炎的炎症事件。MEN 16132是一种高效的B-2受体拮抗剂，能够阻断人滑膜细胞中诱发的BK的促炎反应。
Background and purpose:Bradykinin (BK) and B-2 receptors have been implicated in the pathophysiology of osteoarthritis (OA), and synovitis is one of its hallmarks. Here, the selective B-2 receptor antagonists MEN16132 and icatibant have been pharmacologically characterized in human synovial cells.Experimental approach:Radioligand and functional studies (inositol phosphate (IP) accumulation, interleukin (IL)-6 and IL-8 release) were performed in cultured synoviocytes.Key results:[3H]-BK saturation studies indicated receptor density (B-max) and K-d values of 121 550 sites per cell and 1.14 nM respectively. In synoviocytes, MEN16132 (pK(i) 8.9) was threefold more potent than icatibant (pK(i) 8.4). Both antagonists showed competitive antagonism in the BK-induced IP assay (control EC50 0.45 nM), with pK(B) values of 9.9 (MEN16132) and 8.1 (icatibant). 24h incubation with BK induced IL-6 (EC50 216 nM) and IL-8 (EC50 53 nM) release. Both MEN16132 (IL-6: pIC(50) 8.1; IL-8: pIC(50) 8.4) and icatibant (IL-6: pIC(50) 6.6; IL-8: pIC(50) 6.7) completely prevented this BK-induced release. Indomethacin did not affect the basal or the IL-6/IL-8 release induced by BK, whereas nordihydroguaiaretic acid decreased the basal release, although BK still increased IL-6 and IL-8 production. BK-induced IL-8 release was attenuated by inhibitors of phospholipase C (U73122), p38 (SB203580), JNK (SP600125), ERK 1/2 (PD98059) MAPKs, phosphoinositide 3-kinase (LY294002), NF-kappa b (BAY-117085) and by the glucocorticoid dexamethasone.Conclusions and implications:Bradykinin via B-2 receptors can participate in inflammatory events in synovitis. MEN16132 is a highly potent B-2 receptor antagonist capable of blocking pro-inflammatory responses to BK evoked in human synoviocytes.