The effect of selective phosphodiesterase isoenzyme inhibition on neutrophil function in vitro
The effect of selective phosphodiesterase isoenzyme inhibition on neutrophil function in vitro
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DOI：
10.1016/j.pupt.2004.10.001
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发表时间：
2005-01-01
影响因子：
3.2
通讯作者：
Page, CP
中科院分区：
文献类型：
--
作者：
Jones, NA;Boswell-Smith, V;Page, CP
中性粒细胞衍生的蛋白水解酶，如中性粒细胞弹性蛋白酶(NE)和基质金属蛋白酶(NIMP)参与了慢性阻塞性肺疾病(COPD)的发病过程。本研究观察了选择性磷酸二酯酶(PDE)抑制人脐静脉内皮细胞(HUVECs)释放去甲肾上腺素(NE)、基质金属蛋白酶-9(MMP9)、髓过氧化物酶(MPO)活性及整合素介导中性粒细胞与内皮细胞(HUVECs)黏附的影响。在没有和存在肿瘤坏死因子(肿瘤坏死因子)(100U ml(-1))的情况下，用PDE抑制剂(10(-11)-10(-4)M)处理中性粒细胞30分钟，然后激活fMLP。45min后取出细胞，测定去甲肾上腺素、髓过氧化物酶和基质金属蛋白酶9的释放。在粘附性研究中，中性粒细胞被铬-51放射性标记，刺激并立即转移到培养的HUVEC单层中30min，然后评估粘附性。在MPO活性(P<0.01)和NE释放(P<0.01)方面，肿瘤坏死因子-α(100 U·ml~(-1))与fMLP有协同作用。相反，在基质金属蛋白酶-9的释放和中性粒细胞与人脐静脉内皮细胞的黏附方面，肿瘤坏死因子-α和fMLP具有相加的作用。PDE4抑制剂罗氟司特、罗氟司特N-氧化物、西洛司特和罗力普兰均显著抑制有或无肿瘤坏死因子-α时MPO、NE和基质金属蛋白酶-9的释放(P<；0.05；n=6-10)，并减少中性粒细胞与HUVECs的黏附。相反，米力农，一种PDE3抑制剂和非选择性PDE抑制剂，茶碱在任何实验条件下都不能抑制嗜天青细胞脱颗粒。这些数据进一步证明，选择性PDE4同工酶抑制剂可以抑制中性粒细胞脱颗粒，这是PDE3抑制剂或茶碱所不具备的效果。(C)2004爱思唯尔有限公司。保留所有权利。
Neutrophil-derived proteases such as neutrophil elastase (NE) and matrix metalloproteinase (NIMP) are implicated in the pathogenesis of Chronic Obstructive Pulmonary Disease (COPD). In this study, the effects of selective phosphodiesterase (PDE) inhibition on NE and MMP-9 release, as well as Myeloperoxidase (MPO) activity and integrin-mediated neutrophil adhesion to human umbilical vein endothelial cells (HUVECs), were investigated. Human neutrophils were treated with PDE inhibitors (10(-11) -10(-4) M) in the absence and presence of TNF-alpha (tumour necrosis factor) (100 U ml(-1)) for 30 min, prior to fMLP activation. After 45 min, the cells were removed and NE, MPO and MMP-9 release assessed. In the adhesion studies, the neutrophils were radio-labelled with Cr-51, stimulated and immediately transferred to cultured HUVEC monolayers for 30 min, prior to assessment of adhesion. TNF-alpha (100 U ml(-1)) acted synergistically with fMLP in stimulating azurophil degranulation with respect to both MPO activity (P < 0.01) and NE release (P < 0.01). In contrast, an additive effect was observed with TNF-alpha and fMLP with regard to MMP-9 release and neutrophil adhesion to HUVECs. The PDE4 inhibitors, roflumilast, roflumilast N-oxide, cilomilast and rolipram significantly suppressed MPO, NE and MMP-9 release in both the presence and absence of TNF-alpha (P < 0.05; n = 6-10) and also reduced neutrophil adhesion to HUVECs. In contrast, milrinone, a PDE3 inhibitor and the non-selective PDE inhibitor, theophylline did not inhibit azurophil degranulation under any of the experimental conditions. These data provide further evidence that selective PDE4 isoenzyme inhibitors can inhibit neutrophil degranulation, effects not shared by PDE3 inhibitors or theophylline. (C) 2004 Elsevier Ltd. All rights reserved.