Critical role of COX-1 in prostacyclin production by human endothelial cells under modification of hydroperoxide tone
Critical role of COX-1 in prostacyclin production by human endothelial cells under modification of hydroperoxide tone
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DOI：
10.1096/fj.08-106591
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发表时间：
2009-02-01
期刊：
影响因子：
4.8
通讯作者：
Sala, Angelo
中科院分区：
文献类型：
--
作者：
Bolego, Chiara;Buccellati, Carola;Sala, Angelo
我们的目的是评估在有或没有外源性花生四烯酸和/或改变细胞内氧化还原平衡的静态条件下，环氧合酶(COX)-1和COX-2对内皮细胞合成前列环素的相对贡献。COX-1(SC560和FR122047)或COX-2(SC236)的选择性抑制剂浓度依赖地(1-300 nM)可使基础和白介素1β诱导的人脐静脉内皮细胞产生的前列环素减少70%或更多；全血实验证实化合物的选择性(IC50 COX-1/COX-2：SC-560：13 nM/930 nM；SC-236：9µM/457 nM)。观察到的异前列腺素伴随形成似乎与COX酶活性有关，而COX-1/COX-2异二聚体的形成是通过免疫沉淀检测到的。在外源性或由血小板激活提供的花生四烯酸和12-羟基过氧二十碳四烯酸存在下，或在谷胱甘肽耗竭后，COX-1抑制而不是COX-2抑制浓度依赖地减少前列环素的产生。这两种异构体似乎都有助于内皮细胞产生基础前列环素，环氧合酶-2提供环氧合酶-1活性所需的过氧化氢。相反，在细胞内谷胱甘肽耗竭或花生四烯酸和过氧化氢的可用性增加的情况下，选择性的COX-2抑制并不显著影响内皮前列环素的产生。这些发现有助于更好地理解环氧合酶抑制剂对前列环素产生的影响。-Bolego，C.，Buccellati，C.，Prada，A.，Gaion，R.M.，Folco，G.，Sala，A.环氧合酶-1在过氧化氢张力修饰下人内皮细胞产生前列环素的关键作用。FASE B J.23,605-612(2009)
We aimed at evaluating the relative contribution of cyclooxygenase (COX) -1 and COX-2 to the synthesis of prostacyclin in endothelial cells under static conditions in the presence or absence of exogenous arachidonic acid and/or altered intracellular redox balance. Selective inhibitors of either COX-1 (SC560 and FR122047) or COX-2 (SC236) concentration dependently (1-300 nM) reduced basal and interleukin (IL) -1 beta-induced prostacyclin production in human umbilical vein endothelial cells by 70% or more; compound selectivity was confirmed using a whole-blood assay (IC50 COX-1/COX-2: 13 nM/930 nM for SC-560; 9 mu M/457 nM for SC-236). The observed concomitant formation of isoprostane appeared to be associated with COX enzyme activity, while formation of COX-1/COX-2 heterodimers was detected by immunoprecipitation. In the presence of arachidonic acid and 12-hydroperoxy-eicosatetraenoic acid, either exogenous or provided by platelet activation, or after glutathione depletion, COX-1 inhibition but not COX-2 inhibition concentration dependently decreased prostacyclin production. Both isoforms appear to contribute to basal prostacyclin production by endothelial cells, with COX-2 providing the hydroperoxide tone required for COX-1 activity. Conversely, in the case of intracellular glutathione depletion or enhanced availability of arachidonic acid and hydroperoxides, selective COX-2 inhibition did not significantly affect the production of endothelial prostacyclin. These findings contribute to a better understanding of the effects of cyclooxygenase inhibitors on prostacyclin production.-Bolego, C., Buccellati, C., Prada, A., Gaion, R. M., Folco, G., Sala, A. Critical role of COX-1 in prostacyclin production by human endothelial cells under modification of hydroperoxide tone. FASEB J. 23, 605-612 (2009)