蛇毒三肽pENW的抗血小板黏附作用及其机制研究

发布时间：2018-03-02 05:07

本文关键词： 蛇毒血小板黏附血栓形成　出处：《药学学报》2015年09期 　论文类型：期刊论文

【摘要】：考察蛇毒三肽(p ENW)对血小板与纤维蛋白原黏附的抑制作用及其机制。采用平板法检测p ENW(116.5~466.2μmol·L-1)对血小板与纤维蛋白原黏附的抑制作用以及纤维蛋白凝块回缩的抑制作用。MTT法测定p ENW对血小板活力的影响;双波长荧光分光光度计法测定血小板胞浆[Ca2+]i变化;流式细胞术测定血小板内活性氧族(ROS)含量;ELISA法测定血小板内环磷酸鸟苷(c GMP)、环磷酸腺苷(c AMP)、血栓烷A2(TXA2)的水平;Western blot法测定p ENW对血小板内Akt、ERK与p38磷酸化水平的影响。实验结果显示,p ENW显著抑制血小板与纤维蛋白原的黏附及凝血酶诱导纤维蛋白凝块的回缩;p ENW可升高血小板内c GMP/c AMP含量,抑制TXA2的生成及抑制血小板胞浆内[Ca2+]i升高。p ENW抑制凝血酶诱导血小板内的Akt信号通路、ERK与p38信号通路的磷酸化,p ENW对血小板的抑制作用与ROS无关。结果表明,p ENW能够有效抑制血小板与纤维蛋白原的黏附及纤维蛋白凝块的回缩,因此p ENW既可以阻止血栓形成的始动环节,又能减缓已形成血栓的固化过程。
[Abstract]:To investigate the inhibitory effect of venom tripeptide pENW on platelet adhesion to fibrinogen and its mechanism. The inhibitory effect of p ENW(116.5~466.2 渭 mol 路L -1 on platelet adhesion to fibrinogen and the inhibition of fibrin clot retraction were detected by plate method. The effect of p ENW on platelet activity was measured by. The changes of platelet cytosol [Ca2] I were measured by dual wavelength fluorescence spectrophotometer. Determination of reactive oxygen species in platelets by flow cytometry and Elisa for the determination of intracellular cyclic guanosine monophosphate, cyclic adenosine monophosphate c AMPN, thromboxane A _ 2 TXA2) and the effect of p ENW on the phosphorylation of Akttterk and p38 in platelets by Western blot. The results showed that p ENW significantly inhibited platelet adhesion to fibrinogen and thrombin induced fibrin clot retraction p ENW increased the content of c GMP/c AMP in platelets. The inhibition of TXA2 production and the increase of [Ca2] I in platelets. P ENW inhibited the phosphorylation of Akt signaling pathway and p38 signaling pathway induced by thrombin. The results showed that the inhibitory effect of ENW on platelet was not related to ROS. ENW can effectively inhibit the adhesion of platelet to fibrinogen and the retraction of fibrin clots. Therefore, p ENW can not only prevent the initiation of thrombosis, but also slow down the process of thrombus solidification.
【作者单位】：河南中医学院;中国药科大学药学院;
【基金】：国家自然科学基金资助项目(81173088) 河南省科技攻关重点项目(102102310320) 河南中医学院博士科研基金资助项目(BSJJ2012-01)
【分类号】：R96

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