NO 供体、硝普钠、SIN-1 通过激活 cGMP 非依赖性途径抑制人及大鼠血小板的体外作用。

In vitro inhibition of human and rat platelets by NO donors, nitrosoglutathione, sodium nitroprusside and SIN-1, through activation of cGMP-independent pathways.

机构信息

Department of Neuroscience, Pharmacology Unit, University of Siena, 53100 Siena, Italy.

出版信息

Pharmacol Res. 2011 Sep;64(3):289-97. doi: 10.1016/j.phrs.2011.03.014. Epub 2011 Apr 23.

DOI:10.1016/j.phrs.2011.03.014

Abstract

Three different NO donors, S-nitrosoglutathione (GSNO), sodium nitroprusside (SNP) and 3-morpholino-sydnonimine hydrochloride (SIN-1) were used in order to investigate mechanisms of platelet inhibition through cGMP-dependent and -independent pathways both in human and rat. To this purpose, we also evaluated to what extent cGMP-independent pathways were related with the entity of NO release from each drug. SNP, GSNO and SIN-1 (100 μM) effects on platelet aggregation, in the presence or absence of a soluble guanylate cyclase inhibitor (ODQ), on fibrinogen receptor (α(IIb)β(3)) binding to specific antibody (PAC-1), and on the entity of NO release from NO donors in human and rat platelet rich plasma (PRP) were measured. Inhibition of platelet aggregation (induced by ADP) resulted to be greater in human than in rat. GSNO was the most powerful inhibitor (IC(50) values, μM): (a) in human, GSNO=0.52±0.09, SNP=2.83 ± 0.53, SIN-1=2.98 ± 1.06; (b) in rat, GSNO = 28.4 ± 6.9, SNP = 265 ± 73, SIN-1=108 ± 85. GSNO action in both species was mediated by cGMP-independent mechanisms and characterized by the highest NO release in PRP. SIN-1 and SNP displayed mixed mechanisms of inhibition of platelet aggregation (cGMP-dependent and independent), except for SIN-1 in rat (cGMP-dependent), and respectively lower or nearly absent NO delivery. Conversely, all NO-donors prevalently inhibited PAC-1 binding to α(IIb)β(3) through cGMP-dependent pathways. A modest relationship between NO release from NO donors and cGMP-independent responses was found. Interestingly, the species difference in NO release from GSNO and inhibition by cGMP-independent mechanism was respectively attributed to S-nitrosylation of non-essential and essential protein SH groups.

摘要

三种不同的一氧化氮供体，S-亚硝基谷胱甘肽（GSNO）、硝普钠（SNP）和 3-吗啉-sydnonimine 盐酸盐（SIN-1）被用于研究通过 cGMP 依赖性和非依赖性途径抑制血小板的机制，无论是在人类还是大鼠中。为此，我们还评估了 cGMP 非依赖性途径与每种药物释放的 NO 量之间的关系。在存在或不存在可溶性鸟苷酸环化酶抑制剂（ODQ）的情况下，测量了 SNP、GSNO 和 SIN-1（100μM）对血小板聚集的影响，纤维蛋白原受体（α（IIb）β（3））与特异性抗体（PAC-1）结合，以及在人类和大鼠富含血小板的血浆（PRP）中 NO 供体释放的 NO 量。ADP 诱导的血小板聚集抑制作用在人类中比在大鼠中更强。GSNO 是最强的抑制剂（IC50 值，μM）：（a）在人类中，GSNO=0.52±0.09，SNP=2.83 ± 0.53，SIN-1=2.98 ± 1.06；（b）在大鼠中，GSNO=28.4 ± 6.9，SNP=265 ± 73，SIN-1=108 ± 85。GSNO 在两种物种中的作用均通过 cGMP 非依赖性机制介导，并具有 PRP 中最高的 NO 释放。SIN-1 和 SNP 均显示出抑制血小板聚集的混合机制（cGMP 依赖性和非依赖性），但 SIN-1 在大鼠中为 cGMP 依赖性，并且分别为较低或几乎不存在的 NO 传递。相反，所有的 NO 供体都通过 cGMP 依赖性途径优先抑制 PAC-1 与α（IIb）β（3）的结合。发现 NO 供体从 NO 供体中释放和 cGMP 非依赖性反应之间存在适度的关系。有趣的是，从 GSNO 中释放 NO 和通过 cGMP 非依赖性机制抑制的种间差异分别归因于非必需和必需蛋白 SH 基团的 S-亚硝基化。