Iloprost inhibits superoxide formation and gp91phox expression induced by the thromboxane A2 analogue U46619, 8-isoprostane F2α, prostaglandin F2α, cytokines and endotoxin in the pig pulmonary artery
Iloprost inhibits superoxide formation and gp91phox expression induced by the thromboxane A2 analogue U46619, 8-isoprostane F2α, prostaglandin F2α, cytokines and endotoxin in the pig pulmonary artery
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DOI：
10.1038/sj.bjp.0705626
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发表时间：
2004-02-01
影响因子：
7.3
通讯作者：
Jeremy, JY
中科院分区：
文献类型：
--
作者：
Muzaffar, S;Shukla, N;Jeremy, JY
由于血栓素A(2) （TXA(2)）、前列腺素（PGI(2)）和8-异前列腺素F-2alpha在介导血管O-2（环）形成中的作用及其与成人呼吸窘迫综合征（ARDS）的关系尚不清楚，因此我们研究了这些类20烷对培养猪肺动脉（PA）节段、PA血管平滑肌细胞（PAVSMCs）和PA内皮细胞（PAECs）中gp91(phox) （NADPH氧化酶的催化亚基）表达和O-2（环）释放的影响PA片段、PAVSMCs和paec与TXA(2)类似物U46619、（+/- lps、肿瘤坏死因子- α (tnf - α)或il -1 α）、8-异前列腺素f -2 α和+/- iloprost（稳定的PGI(2)类似物）一起培养。然后用分光光度法测定超氧化物歧化酶抑制的O-2（环-）的形成，用Western blotting测定gp91（phox）的表达。在平行实验中，用LPS、tnf - α和il - α处理整个PA段，然后用酶联免疫分析法测定TXA(2)、PGI(2)、PGF（2α）和8-异前列腺素f - 2α的形成U46619、PGF（2alpha）和8-异前列腺素F-2alpha可促进PA段、PAVSMCs和PAECs中O-2（环-）的形成，该作用被NADPH氧化酶抑制剂二苯乙烯多胺和夹心草碱抑制，上调PAECs和PAVSMCs中gp91（phox）的表达。脂多糖、tnf - α和il -1 α增强了这些作用，但伊洛前列素抑制了这些作用。在相同的培养条件下，il -1 α、LPS和tnf - α均诱导TXA(2)、PGF（2 α）和8-异前列腺素f -2 α的形成增加，但减少伴随的PGI的形成(2)4这些数据表明，LPS和细胞因子影响猪PA中TXA(2)、PGI(2)、PGF（2alpha）和8-异前列腺素F-2alpha的相对平衡，从而改变NADPH氧化酶的表达和O-2（圆-）的形成。这些新发现对制定治疗ARDS的有效策略具有重要意义。
I Since the roles of thromboxane A(2) (TXA(2)), prostacyclin (PGI(2)) and 8-isoprostane F-2alpha in mediating vascular O-2(circle-) formation and its relation to adult respiratory distress syndrome (ARDS) is unknown, the effects of these eicosanoids on the expression of gp91(phox) (catalytic subunit of NADPH oxidase) and O-2(circle-) release from cultured pig pulmonary artery (PA) segments, PA vascular smooth muscle cells (PAVSMCs) and PA endothelial cells (PAECs) were investigated.2 PA segments, PAVSMCs and PAECs were incubated with the TXA(2) analogue, U46619, (+/-LPS, tumour necrosing factor-alpha (TNF-alpha) or IL-1alpha), 8-isoprostane F-2alpha and +/- iloprost (a stable PGI(2) analogue) for 16 It. The formation of superoxide dismutase-inhibitable O-2(circle-) was then measured spectrophotometrically and gp91(phox) expression assessed using Western blotting. In parallel experiments, whole PA segments were treated with LPS, TNF-alpha and IL-alpha after which time TXA(2), PGI(2), PGF(2alpha) and 8-isoprostane F-2alpha formation was measured using enzyme-linked immunoassays.3 U46619, PGF(2alpha) and 8-isoprostane F-2alpha promoted the formation of O-2(circle-) in PA segments, PAVSMCs and PAECs, an effect inhibited by diphenyleneiodonium and apocynin (both NADPH oxidase inhibitors) and upregulated the expression of gp91(phox) in PAECs and PAVSMCs. These effects were augmented by LPS, TNF-alpha and IL-1alpha but inhibited by iloprost. Under identical incubation conditions, IL-1alpha, LPS and TNF-alpha all induced an increase in the formation of TXA(2), PGF(2alpha) and 8-isoprostane F-2alpha but reduced the concomitant formation of PGI(2).4 These data demonstrate that LPS and cytokines influence the relative balance of TXA(2), PGI(2), PGF(2alpha) and 8-isoprostane F-2alpha in pig PA, which in turn alter NADPH oxidase expression and O-2(circle-) formation. These novel findings have implications in devising effective strategies for treating ARDS.