Combined Blockade of ADP Receptors and PI3-Kinase p110β Fully Prevents Platelet and Leukocyte Activation during Hypothermic Extracorporeal Circulation
Combined Blockade of ADP Receptors and PI3-Kinase p110β Fully Prevents Platelet and Leukocyte Activation during Hypothermic Extracorporeal Circulation
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DOI：
10.1371/journal.pone.0038455
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发表时间：
2012-06-06
期刊：
影响因子：
3.7
通讯作者：
Straub, Andreas
中科院分区：
文献类型：
--
作者：
Krajewski, Stefanie;Kurz, Julia;Straub, Andreas
体外循环（ECC）和低温是心脏手术中维持稳定循环参数和提高患者缺血耐受性的重要手段。然而，ECC和低温诱导血小板和白细胞的活化机制，这是由血小板激动剂ADP和磷酸肌醇-3-激酶（PI 3 K）p110 β介导的。在临床条件下，这些过程与危及生命的并发症相关，包括血栓栓塞和炎症。本研究分析了ADP受体P2 Y12和P2 Y1阻断和PI 3 K p110 β抑制对低温ECC期间血小板和粒细胞的影响。用P2 Y12拮抗剂2-MeSAMP、P2 Y1拮抗剂MRS 2179、PI 3 K p110 β抑制剂TGX-221、其组合或PBS和丙二醇（对照）处理人血液。在静态体外条件下，使用2-MeSAMP或TGX-221发现了关于ADP诱导的血小板活化抑制的浓度依赖性效应。用MRS 2179实现了ADP介导的作用的进一步抑制。接下来，在28 ℃下使血液在离体ECC模型中循环30分钟，并使用流式细胞术、ELISA和血小板计数分析研究各种血小板和粒细胞标志物。单独使用TGX-221或与P2 Y阻断剂联合使用可抑制低温ECC诱导的GPIIb/IIIa激活（p
Extracorporeal circulation (ECC) and hypothermia are used to maintain stable circulatory parameters and improve the ischemia tolerance of patients in cardiac surgery. However, ECC and hypothermia induce activation mechanisms in platelets and leukocytes, which are mediated by the platelet agonist ADP and the phosphoinositide-3-kinase (PI3K) p110 beta. Under clinical conditions these processes are associated with life-threatening complications including thromboembolism and inflammation. This study analyzes effects of ADP receptor P2Y12 and P2Y1 blockade and PI3K p110 beta inhibition on platelets and granulocytes during hypothermic ECC. Human blood was treated with the P2Y12 antagonist 2-MeSAMP, the P2Y1 antagonist MRS2179, the PI3K p110 beta inhibitor TGX-221, combinations thereof, or PBS and propylene glycol (controls). Under static in vitro conditions a concentration-dependent effect regarding the inhibition of ADP-induced platelet activation was found using 2-MeSAMP or TGX-221. Further inhibition of ADP-mediated effects was achieved with MRS2179. Next, blood was circulated in an ex vivo ECC model at 28 degrees C for 30 minutes and various platelet and granulocyte markers were investigated using flow cytometry, ELISA and platelet count analysis. GPIIb/IIIa activation induced by hypothermic ECC was inhibited using TGX-221 alone or in combination with P2Y blockers (p