Modrego Javier, Azcona Luis, Martín-Palacios Naiara, Zamorano-León José J, Segura Antonio, Rodríguez Pablo, Guerra Reddy, Tamargo Juan, Macaya Carlos, López-Farré Antonio J
Cardiovascular Research Unit, Hospital Clínico San Carlos, Universidad Complutense de Madrid, Madrid, Spain.
Cardiovascular Research Unit, Hospital Clínico San Carlos, Universidad Complutense de Madrid, Madrid, Spain ; Hemodynamic Unit, Cardiology Department, Hospital Clínico San Carlos, Universidad Complutense de Madrid, Madrid, Spain.
PLoS One. 2013 Dec 20;8(12):e82574. doi: 10.1371/journal.pone.0082574. eCollection 2013.
To analyse if platelet responsiveness to aspirin (ASA) may be associated with a different ability of platelets to generate nitric oxide (NO).
PATIENTS/METHODS: Platelets were obtained from 50 patients with stable coronary ischemia and were divided into ASA-sensitive (n = 26) and ASA-resistant (n = 24) using a platelet functionality test (PFA-100).
ASA-sensitive platelets tended to release more NO (determined as nitrite + nitrate) than ASA-resistant platelets but it did not reach statistical significance. Protein expression of nitric oxide synthase 3 (NOS3) was higher in ASA-sensitive than in ASA-resistant platelets but there were no differences in the platelet expression of nitric oxide synthase 2 (NOS2) isoform. The highest NOS3 expression in ASA-sensitive platelets was independent of the presence of T-to-C mutation at nucleotide position -786 (T(-786) → C) in the NOS3-coding gene. However, platelet content of phosphorylated NOS3 at Serine (Ser)(1177), an active form of NOS3, was higher in ASA-sensitive than in ASA-resistant platelets. The level of platelet NOS3 Ser(1177) phosphorylation was positively associated with the closure time in the PFA-100 test. In vitro, collagen failed to stimulate the aggregation of ASA-sensitive platelets, determined by lumiaggregometry, and it was associated with a significant increase (p = 0.018) of NOS3 phosphorylation at Ser(1177). On the contrary, collagen stimulated the aggregation of ASA-resistant platelets but did not significantly modify the platelet content of phosphorylated NOS3 Ser(1177). During collagen stimulation the release of NO from ASA-sensitive platelets was significantly enhanced but it was not modified in ASA-resistant platelets.
Functional platelet responsiveness to ASA was associated with the platelet content of phosphorylated NOS3 at Ser(1177).
分析血小板对阿司匹林（ASA）的反应性是否可能与血小板产生一氧化氮（NO）的不同能力相关。
患者/方法：从50例稳定型冠状动脉缺血患者中获取血小板，并使用血小板功能测试（PFA-100）将其分为ASA敏感组（n = 26）和ASA抵抗组（n = 24）。
ASA敏感血小板比ASA抵抗血小板倾向于释放更多的NO（以亚硝酸盐+硝酸盐测定），但未达到统计学意义。一氧化氮合酶3（NOS3）的蛋白表达在ASA敏感血小板中高于ASA抵抗血小板，但一氧化氮合酶2（NOS2）亚型的血小板表达无差异。ASA敏感血小板中最高的NOS3表达与NOS3编码基因核苷酸位置-786（T（-786）→C）处T到C突变的存在无关。然而，一氧化氮合酶3的活性形式——丝氨酸（Ser）（1177）处磷酸化的NOS3的血小板含量在ASA敏感血小板中高于ASA抵抗血小板。血小板NOS3 Ser（1177）磷酸化水平与PFA-100测试中的封闭时间呈正相关。在体外，通过光聚集测定法确定，胶原蛋白未能刺激ASA敏感血小板的聚集，并且这与Ser（1177）处NOS3磷酸化的显著增加（p = 0.018）相关。相反，胶原蛋白刺激了ASA抵抗血小板的聚集，但未显著改变磷酸化的NOS3 Ser（1177）的血小板含量。在胶原蛋白刺激期间，ASA敏感血小板中NO的释放显著增强，但ASA抵抗血小板中未改变。
血小板对ASA的功能性反应性与Ser（1177）处磷酸化的NOS3的血小板含量相关。
