Laboratory of Veterinary Physiology and Cell Signaling, College of Veterinary Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.
Evid Based Complement Alternat Med. 2013;2013:569160. doi: 10.1155/2013/569160. Epub 2013 Aug 13.
A number of reagents that prevent thrombosis have been developed but were found to have serious side effects. Therefore, we sought to identify complementary and alternative medicinal materials that are safe and have long-term efficacy. In the present studies, we have assessed the ability of chlorine e6 (CE6) to inhibit ADP-induced aggregation of rat platelets and elucidated the underlying mechanism. CE6 inhibited platelet aggregation induced by 10 µM ADP in a concentration-dependent manner and decreased intracellular calcium mobilization and granule secretion (i.e., ATP and serotonin release). Western blotting revealed that CE6 strongly inhibited the phosphorylations of PI3K, Akt, c-Jun N-terminal kinase (JNK), and different mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase 1/2 (ERK1/2) as well as p38-MAPK. Our study also demonstrated that CE6 significantly elevated intracellular cAMP levels and decreased thromboxane A2 formation in a concentration-dependent manner. Furthermore, we determined that CE6 initiated the activation of PKA, an effector of cAMP. Taken together, our findings indicate that CE6 may inhibit ADP-induced platelet activation by elevating cAMP levels and suppressing PI3K/Akt activity. Finally, these results suggest that CE6 could be developed as therapeutic agent that helps prevent thrombosis and ischemia.
已经开发出了许多防止血栓形成的试剂，但发现它们有严重的副作用。因此，我们试图寻找安全且具有长期疗效的补充和替代药材。在本研究中，我们评估了氯乙啶 6（CE6）抑制 ADP 诱导的大鼠血小板聚集的能力，并阐明了其潜在机制。CE6 以浓度依赖性方式抑制由 10 μM ADP 诱导的血小板聚集，并减少细胞内钙动员和颗粒分泌（即 ATP 和 5-羟色胺释放）。Western blot 显示 CE6 强烈抑制 PI3K、Akt、c-Jun N 端激酶（JNK）和不同丝裂原活化蛋白激酶（MAPK）（包括细胞外信号调节激酶 1/2（ERK1/2））以及 p38-MAPK 的磷酸化。我们的研究还表明，CE6 可显著升高细胞内 cAMP 水平，并以浓度依赖性方式降低血栓烷 A2 的形成。此外，我们确定 CE6 启动了 cAMP 的效应物 PKA 的激活。综上所述，我们的研究结果表明，CE6 可能通过升高 cAMP 水平和抑制 PI3K/Akt 活性来抑制 ADP 诱导的血小板激活。最后，这些结果表明，CE6 可开发为有助于预防血栓形成和缺血的治疗剂。
