葡萄籽原花青素提取物对体内大鼠再灌注心律失常的保护作用的分子机制。

The molecular mechanism of protective effects of grape seed proanthocyanidin extract on reperfusion arrhythmias in rats in vivo.

机构信息

Department of Geriatrics, Shandong University Qilu Hospital, Jinan 250012, China.

出版信息

Biol Pharm Bull. 2010;33(5):759-67. doi: 10.1248/bpb.33.759.

DOI:10.1248/bpb.33.759

PMID:20460751

Abstract

Reperfusion arrhythmias (RA) especially ventricular tachycardia (VT) and ventricular fibrillation (VF) remain the most important causes of sudden death following reperfusion. In isolated rat hearts grape seed proanthocyanidin extract (GSPE) had been proved to reduce the incidence of reperfusion-induced VF and VT. However the mechanism of this protection remained unclear. The aim of this study was to elucidate the potential mechanism of this protection of GSPE. The myocardial ischemia reperfusion (IR) model was induced by 30 min coronary occlusion and 120 min reperfusion in open chest anesthetized rats. The ultrastructure of ischemic cardiomyocytes was observed. An isobaric tag labeling for relative and absolute quantification (iTRAQ) proteomics was used to identify differentially expressed membrane proteins. Western blot was performed to verify the results of iTRAQ. The results demonstrated GSPE can significantly reduce the incidence of VT and VF induced by reperfusion in vivo. We identified 92 differentially expressed proteins. Western blot analysis confirmed GSPE increased the expression of Na(+)/K(+)-ATPase alpha1 subunit (p<0.01). We found the subunit distribution of Na(+)/K(+)-ATPase was changed after reperfusion. Na(+)/K(+)-ATPase alpha1 subunit was decreased in IR group (without GSPE-treated) compared to sham group while it was significantly increased in GSPE group. The decrease of free radical generation induced by GSPE may lead to the up-regulation of Na(+)/K(+)-ATPase alpha1 subunit. This change of subunit distribution may lead to the increase of activity of Na(+)/K(+)-ATPase which may result in the protection of GSPE against reperfusion arrhythmias. Our experiments provided new avenues for the treatment of reperfusion arrhythmias.

摘要

再灌注心律失常（RA），特别是室性心动过速（VT）和心室颤动（VF），仍然是再灌注后猝死的最重要原因。在分离的大鼠心脏中，已证实葡萄籽原花青素提取物（GSPE）可降低再灌注诱导的VF 和 VT 的发生率。然而，这种保护的机制尚不清楚。本研究旨在阐明 GSPE 这种保护作用的潜在机制。采用开胸麻醉大鼠 30 分钟冠状动脉闭塞和 120 分钟再灌注的方法诱导心肌缺血再灌注（IR）模型。观察缺血性心肌细胞的超微结构。采用等压标签标记相对和绝对定量（iTRAQ）蛋白质组学方法鉴定差异表达的膜蛋白。采用 Western blot 验证 iTRAQ 的结果。结果表明，GSPE 可显著降低体内再灌注诱导 VT 和 VF 的发生率。我们鉴定出 92 种差异表达蛋白。Western blot 分析证实 GSPE 可增加 Na(+)/K(+)-ATPase alpha1 亚基的表达（p<0.01）。我们发现再灌注后 Na(+)/K(+)-ATPase 的亚基分布发生了变化。与 sham 组相比，IR 组（未用 GSPE 处理）Na(+)/K(+)-ATPase alpha1 亚基减少，而 GSPE 组明显增加。GSPE 减少自由基生成可能导致 Na(+)/K(+)-ATPase alpha1 亚基上调。这种亚基分布的变化可能导致 Na(+)/K(+)-ATPase 活性增加，从而对 GSPE 防治再灌注心律失常起到保护作用。我们的实验为再灌注心律失常的治疗提供了新的途径。