Adenosine inhibits matrix metalloproteinase-9 secretion by neutrophils -: Implication of A2a receptor and cAMP/PKA/Ca2+ pathway
Adenosine inhibits matrix metalloproteinase-9 secretion by neutrophils -: Implication of A2a receptor and cAMP/PKA/Ca2+ pathway
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DOI：
10.1161/01.res.0000241428.82502.d4
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发表时间：
2006-09-15
影响因子：
20.1
通讯作者：
Wagner, Daniel R.
中科院分区：
文献类型：
--
作者：
Ernens, Isabelle;Rouy, Didier;Wagner, Daniel R.
基质金属蛋白酶（MMPs），特别是中性粒细胞分泌的MMP-9，能够降解心脏的基质成分，被认为是梗死后心肌基质重塑的驱动力。腺苷是一种自然产生的核苷，已被证明具有保护心脏的作用，并抑制各种细胞因子的分泌。我们研究的目的是确定腺苷对中性粒细胞分泌MMP-9的影响。通过Ficoll和右旋糖酐沉淀从健康志愿者中分离中性粒细胞。中性粒细胞在存在或不存在腺苷或腺苷类似物的情况下被n -甲酰基蛋氨酸-亮氨酸-苯丙氨酸（fMLP）激活。酶谱法和酶联免疫吸附法检测MMP-9的分泌。腺苷（1 μ mol/L）使fmlp诱导的MMP-9分泌减少30 +/- 2% （n = 8, P < 0.001）。这种效应是剂量依赖性的，并不针对fMLP，因为腺苷也能抑制LPS或h2o2刺激的中性粒细胞分泌MMP-9。A2a受体激动剂CGS21680能模拟腺苷的作用，A2a拮抗剂SCH5826和A2a RNA沉默均能抑制腺苷的作用。A3激动剂IB-MECA适度降低fmlp诱导的MMP-9分泌。其他类型腺苷受体的激动剂和拮抗剂无显著作用。腺苷增加细胞内cAMP浓度，加速胞浆内钙峰值回归基线。腺苷对MMP-9分泌的抑制作用以及钙效应被蛋白激酶A抑制剂H-89所阻止。总之，我们在这里表明腺苷抑制中性粒细胞分泌MMP-9。我们的研究结果表明，这种作用暗示了A2a受体，并通过cAMP/PKA/Ca2+途径介导。因此，腺苷可能是防止心肌损伤后基质降解和重构的新途径。
Matrix metalloproteinases (MMPs), and in particular MMP-9 secreted by neutrophils, are capable of degrading the matrix components of the heart and are thought to be the driving force behind myocardial matrix remodeling after infarction. Adenosine, a naturally produced nucleoside, has been shown to have cardioprotective effects and to inhibit secretion of various cytokines. The aim of our study was to determine the effect of adenosine on the secretion of MMP-9 by neutrophils. Neutrophils were isolated from healthy volunteers through Ficoll and Dextran sedimentation. Neutrophils were activated by N-formylmethionyl-leucyl-phenylalanine (fMLP) in the presence or absence of adenosine or adenosine analogs. Zymography and enzyme linked immunosorbent assay were used to measure MMP-9 secretion. Adenosine (1 mu mol/L) decreased the fMLP-induced MMP-9 secretion by 30 +/- 2% (n = 8, P < 0.001). The effect was dose-dependent and was not specific to fMLP because adenosine also inhibited MMP-9 secretion by LPS- or H2O2-stimulated neutrophils. The effect of adenosine was mimicked by the adenosine A2a receptor agonist CGS21680 and was inhibited by both the A2a antagonist SCH5826 and A2a RNA silencing. The A3 agonist IB-MECA moderately decreased fMLP-induced MMP-9 secretion. Agonists and antagonists of the other types of adenosine receptors had no significant effect. Adenosine increased intracellular cAMP concentration and accelerated the return to baseline of the intracytoplasmic calcium peak. The inhibition of MMP-9 secretion by adenosine, as well as the calcium effect, was prevented by the protein kinase A inhibitor H-89. In conclusion, we show here that adenosine inhibits MMP-9 secretion by neutrophils. Our results suggest that this effect implies the A2a receptor and is mediated through the cAMP/PKA/Ca2+ pathway. Therefore, adenosine may represent a new approach to prevent matrix degradation and remodeling after myocardial injury.