A novel antithrombotic effect of sulforaphane via activation of platelet adenylate cyclase: ex vivo and in vivo studies
A novel antithrombotic effect of sulforaphane via activation of platelet adenylate cyclase: ex vivo and in vivo studies
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DOI：
10.1016/j.jnutbio.2012.08.007
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发表时间：
2013-06-01
影响因子：
5.6
通讯作者：
Hsieh, Cheng-Ying
中科院分区：
文献类型：
--
作者：
Jayakumar, Thanasekaran;Chen, Wei-Fan;Hsieh, Cheng-Ying
萝卜硫素是一种天然存在的异硫氰酸盐，存在于西兰花和卷心菜等十字花科蔬菜中。研究发现萝卜硫素通过调节多种机制对肿瘤生长具有非常有效的抑制作用。然而，没有关于萝卜硫素对血小板活化的影响及其相关问题的数据。动脉血栓形成引起的血小板活化与多种心血管疾病有关。因此，本研究的目的是检查萝卜硫素的体内抗血栓作用及其在血小板活化中的可能机制。萝卜硫素（0.125 和 0.25 mg/kg）可有效降低 ADP 诱导的小鼠急性肺血栓栓塞的死亡率。其他体内研究还表明，萝卜硫素（0.25 mg/kg）可显着延长小鼠血小板栓塞的形成。此外，萝卜硫素（15-75μM）表现出更有效的抑制胶原蛋白刺激的血小板聚集的活性。萝卜硫素抑制血小板活化，同时抑制相对 Ca2+ 动员；磷脂酶 C (PLC)gamma 2、蛋白激酶 C (PKC)、丝裂原激活蛋白激酶 (MAPK) 和 Akt 的磷酸化；和羟基自由基（OH中心点）的形成。萝卜硫素显着增加环 (c)AMP 水平，但不增加环 (c)GMP 水平，并刺激血管舒张刺激磷蛋白 (VASP) 磷酸化。 SQ22536 是一种腺苷酸环化酶抑制剂，但不能明显逆转萝卜硫素介导的血小板聚集作用，而 ODQ（1H-[1,2,4]Oxadiazolo[4,3-a]quinoxal in-1-one）是一种鸟苷酸环化酶抑制剂。 PKC 激活、p38 MAPK、Akt 和 VASP 磷酸化；和OH中心点的形成。此外，PI3 激酶抑制剂 (LY294002) 和 p38 MAPK 抑制剂 (SB203580) 均显着减少 PKC 激活以及 p38 MAPK 和 Akt 磷酸化；相反，PKC 抑制剂 (RO318220) 不会减弱胶原蛋白刺激的 p38 MAPK 或 Akt 磷酸化。这项研究首次证明，萝卜硫素除了最初被认为是一种预防肿瘤生长的药物外，还具有有效的抗血小板活性，最初可能会激活腺苷酸环化酶/cAMP，然后抑制细胞内信号（例如 PI3-激酶/Akt 和 PLC gamma 2-PKC-p47 级联）并最终抑制血小板活化。因此，萝卜硫素的这种新作用可能代表着治疗或预防心血管疾病的高治疗潜力。 (C) 2013 Elsevier Inc. 保留所有权利。
Sulforaphane is a naturally occurring isothiocyanate, which can be found in cruciferous vegetables such as broccoli and cabbage. Sulforaphane was found to have very potent inhibitory effects on tumor growth through regulation of diverse mechanisms. However, no data are available concerning the effects of sulforaphane on platelet activation and its relative issues. Activation of platelets caused by arterial thrombosis is relevant to a variety of cardiovascular diseases. Hence, the aim of this study was to examine the in vivo antithrombotic effects of sulforaphane and its possible mechanisms in platelet activation. Sulforaphane (0.125 and 0.25 mg/kg) was effective in reducing the mortality of ADP-induced acute pulmonary thromboembolism in mice. Other in vivo studies also revealed that sulforaphane (0.25 mg/kg) significantly prolonged platelet plug formation in mice. In addition, sulforaphane (15-75 mu M) exhibited more-potent activity of inhibiting platelet aggregation stimulated by collagen. Sulforaphane inhibited platelet activation accompanied by inhibiting relative Ca2+ mobilization; phosphorylation of phospholipase C (PLC)gamma 2, protein kinase C (PKC), mitogen-activated protein kinases (MAPKs) and Akt; and hydroxyl radical (OH center dot) formation. Sulforaphane markedly increased cyclic (c)AMP, but not cyclic (c)GMP levels, and stimulated vasodilator-stimulated phosphoprotein (VASP) phosphorylation. SQ22536, an inhibitor of adenylate cyclase, but not ODQ (1H-[1,2,4]Oxadiazolo[4,3-a]quinoxal in-1-one), an inhibitor of guanylate cyclase, obviously reversed the sulforaphane-mediated effects on platelet aggregation; PKC activation, p38 MAPK, Akt and VASP phosphorylation; and OH center dot formation. Furthermore, a PI3-kinase inhibitor (LY294002) and a p38 MAPK inhibitor (SB203580) both significantly diminished PKC activation and p38 MAPK and Akt phosphorylation; in contrast, a PKC inhibitor (RO318220) did not diminish p38 MAPK or Akt phosphorylation stimulated by collagen. This study demonstrates for the first time that in addition to it originally being considered as an agent for prevention of tumor growth, sulforaphane possesses potent antiplatelet activity which may initially activate adenylate cyclase/cAMP, followed by inhibiting intracellular signals (such as the PI3-kinase/Akt and PLC gamma 2-PKC-p47 cascades) and ultimately inhibiting platelet activation. Therefore, this novel role of sulforaphane may represent a high therapeutic potential for treatment or prevention of cardiovascular diseases. (C) 2013 Elsevier Inc. All rights reserved.