Dorothy M. Davis Heart and Lung Research Institute, Division of Cardiovascular Medicine.
Am J Physiol Heart Circ Physiol. 2013 Jan 15;304(2):H294-302. doi: 10.1152/ajpheart.00367.2012. Epub 2012 Nov 16.
The GTP-binding protein Rac regulates diverse cellular functions including activation of NADPH oxidase, a major source of superoxide production (O(2)(·-)). Rac1-mediated NADPH oxidase activation is increased after myocardial infarction (MI) and heart failure both in animals and humans; however, the impact of increased myocardial Rac on impending ischemia-reperfusion (I/R) is unknown. A novel transgenic mouse model with cardiac-specific overexpression of constitutively active mutant form of Zea maize Rac D (ZmRacD) gene has been reported with increased myocardial Rac-GTPase activity and O(2)(·-) generation. The goal of the present study was to determine signaling pathways related to increased myocardial ZmRacD and to what extent hearts with increased ZmRacD proteins are susceptible to I/R injury. The effect of myocardial I/R was examined in young adult wild-type (WT) and ZmRacD transgenic (TG) mice. In vitro reversible myocardial I/R for postischemic cardiac function and in vivo regional myocardial I/R for MI were performed. Following 20-min global ischemia and 45-min reperfusion, postischemic cardiac contractile function and heart rate were significantly reduced in TG hearts compared with WT hearts. Importantly, acute regional myocardial I/R (30-min ischemia and 24-h reperfusion) caused significantly larger MI in TG mice compared with WT mice. Western blot analysis of cardiac homogenates revealed that increased myocardial ZmRacD gene expression is associated with concomitant increased levels of NADPH oxidase subunit gp91(phox), O(2)(·-), and P(21)-activated kinase. Thus these findings provide direct evidence that increased levels of active myocardial Rac renders the heart susceptible to increased postischemic contractile dysfunction and MI following acute I/R.
GTP 结合蛋白 Rac 调节多种细胞功能,包括 NADPH 氧化酶的激活,这是超氧产生(O(2)(·-))的主要来源。在动物和人类中,心肌梗塞(MI)和心力衰竭后 Rac1 介导的 NADPH 氧化酶激活增加;然而,增加的心肌 Rac 对即将发生的缺血再灌注(I/R)的影响尚不清楚。已经报道了一种新型的转基因小鼠模型,其心脏特异性过表达组成型激活的 Zea 玉米 Rac D(ZmRacD)基因,具有增加的心肌 Rac-GTPase 活性和 O(2)(·-)生成。本研究的目的是确定与增加的心肌 ZmRacD 相关的信号通路,以及增加的 ZmRacD 蛋白的心脏对 I/R 损伤的敏感程度。在年轻成年野生型(WT)和 ZmRacD 转基因(TG)小鼠中检查了心肌 I/R 的影响。进行了体外可逆性心肌 I/R 以评估缺血后心脏功能,以及体内区域性心肌 I/R 以评估 MI。在 20 分钟的全缺血和 45 分钟的再灌注后,与 WT 心脏相比,TG 心脏的缺血后心脏收缩功能和心率明显降低。重要的是,急性区域性心肌 I/R(30 分钟缺血和 24 小时再灌注)导致 TG 小鼠的 MI 明显大于 WT 小鼠。心脏匀浆的 Western blot 分析显示,增加的心肌 ZmRacD 基因表达与 NADPH 氧化酶亚基 gp91(phox)、O(2)(·-)和 P21 激活的激酶的水平同时增加有关。因此,这些发现提供了直接证据,表明增加的心肌 Rac 水平使心脏容易发生缺血后收缩功能障碍增加和急性 I/R 后 MI。