Cabigas E Bernadette, Liu Jie, Boopathy Archana V, Che Pao Lin, Crawford Brian H, Baroi Gitangali, Bhutani Srishti, Shen Ming, Wagner Mary B, Davis Michael E
Wallace H. Coulter Department of Biomedical Engineering at Emory University and Georgia Institute of Technology, Atlanta, GA, USA Division of Cardiology, Emory University School of Medicine, Atlanta, GA, USA.
Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, USA Emory+Children's Center for Cardiovascular Biology, Children's Healthcare of Atlanta, Atlanta, GA, USA.
J Cardiovasc Pharmacol Ther. 2015 Jan;20(1):93-103. doi: 10.1177/1074248414533746. Epub 2014 May 15.
In the adult heart, catalase (CAT) activity increases appropriately with increasing levels of hydrogen peroxide, conferring cardioprotection. This mechanism is absent in the newborn for unknown reasons. In the present study, we examined how the posttranslational modification of CAT contributes to its activation during hypoxia/ischemia and the role of c-Abl tyrosine kinase in this process. Hypoxia studies were carried out using primary cardiomyocytes from adult (>8 weeks) and newborn rats. Following hypoxia, the ratio of phosphorylated to total CAT and c-Abl in isolated newborn rat myocytes did not increase and were significantly lower (1.3- and 4.2-fold, respectively; P < .05) than their adult counterparts. Similarly, there was a significant association (P < .0005) between c-Abl and CAT in adult cells following hypoxia (30.9 ± 8.2 to 70.7 ± 13.1 au) that was absent in newborn myocytes. Although ubiquitination of CAT was higher in newborns compared to adults following hypoxia, inhibition of this did not improve CAT activity. When a c-Abl activator (5-(1,3-diaryl-1H-pyrazol-4-yl)hydantoin [DPH], 200 µmol/L) was administered prior to hypoxia, not only CAT activity was significantly increased (P < .05) but also phosphorylation levels were also significantly improved (P < .01) in these newborn myocytes. Additionally, ischemia-reperfusion (IR) studies were performed using newborn (4-5 days) rabbit hearts perfused in a Langendorff method. The DPH given as an intracardiac injection into the right ventricle of newborn rabbit resulted in a significant improvement (P < .002) in the recovery of developed pressure after IR, a key indicator of cardiac function (from 74.6% ± 6.6% to 118.7% ± 10.9%). In addition, CAT activity was increased 3.92-fold (P < .02) in the same DPH-treated hearts. Addition of DPH to adult rabbits in contrast had no significant effect (from 71.3% ± 10.7% to 59.4% ± 12.1%). Therefore, in the newborn, decreased phosphorylation of CAT by c-Abl potentially mediates IR-induced dysfunction, and activation of c-Abl may be a strategy to prevent ischemic injury associated with surgical procedures.
在成年心脏中,过氧化氢酶(CAT)的活性会随着过氧化氢水平的升高而适当增加,从而起到心脏保护作用。但新生儿的这种机制却不明原因地缺失。在本研究中,我们探究了CAT的翻译后修饰如何在缺氧/缺血过程中促进其激活,以及c-Abl酪氨酸激酶在此过程中的作用。使用成年(>8周)和新生大鼠的原代心肌细胞进行缺氧研究。缺氧后,分离的新生大鼠心肌细胞中磷酸化CAT与总CAT以及c-Abl的比值没有增加,且显著低于成年大鼠心肌细胞(分别低1.3倍和4.2倍;P<.05)。同样,缺氧后成年细胞中c-Abl与CAT之间存在显著关联(P<.0005)(从30.9±8.2至70.7±13.1任意单位),而新生心肌细胞中不存在这种关联。尽管缺氧后新生儿中CAT的泛素化水平高于成年人,但抑制这种泛素化并不能提高CAT活性。在缺氧前给予c-Abl激活剂(5-(1,3-二芳基-1H-吡唑-4-基)乙内酰脲[DPH],200µmol/L),这些新生心肌细胞中不仅CAT活性显著增加(P<.05),而且磷酸化水平也显著提高(P<.01)。此外,使用Langendorff方法灌注新生(4-5天)兔心脏进行缺血再灌注(IR)研究。向新生兔右心室内注射心内注射DPH可使IR后心脏功能的关键指标——左心室舒张末压恢复情况显著改善(P<.002)(从74.6%±6.6%提高到118.7%±10.9%)。此外,在相同的DPH处理心脏中,CAT活性增加了3.92倍(P<.02)。相比之下,向成年兔添加DPH没有显著影响(从71.3%±10.7%降至59.4%±12.1%)。因此,在新生儿中,c-Abl介导的CAT磷酸化减少可能介导了IR诱导的功能障碍,激活c-Abl可能是预防与手术相关的缺血性损伤的一种策略。