Centro de Investigaciones Cardiovasculares ¨Dr Horacio E Cingolani¨, CCT-CONICET, Facultad de Ciencias Médicas, Universidad Nacional de La Plata, La Plata, Buenos Aires, Argentina.
Department of Medicine, Pulmonary and Critical Care Medicine, VA Puget Sound Health Care System, University of Washington, Seattle, Washington, USA.
Physiol Rep. 2021 Nov;9(22):e15093. doi: 10.14814/phy2.15093.
We have previously demonstrated that inhibition of extracellularly oriented carbonic anhydrase (CA) isoforms protects the myocardium against ischemia-reperfusion injury. In this study, our aim was to assess the possible further contribution of CA intracellular isoforms examining the actions of the highly diffusible cell membrane permeant inhibitor of CA, ethoxzolamide (ETZ). Isolated rat hearts, after 20 min of stabilization, were assigned to the following groups: (1) Nonischemic control: 90 min of perfusion; (2) Ischemic control: 30 min of global ischemia and 60 min of reperfusion (R); and (3) ETZ: ETZ at a concentration of 100 μM was administered for 10 min before the onset of ischemia and then during the first 10 min of reperfusion. In additional groups, ETZ was administered in the presence of SB202190 (SB, a p38MAPK inhibitor) or chelerythrine (Chel, a protein kinase C [PKC] inhibitor). Infarct size, myocardial function, and the expression of phosphorylated forms of p38MAPK, PKCε, HSP27, and Drp1, and calcineurin Aβ content were assessed. In isolated mitochondria, the Ca response, Ca retention capacity, and membrane potential were measured. ETZ decreased infarct size by 60%, improved postischemic recovery of myocardial contractile and diastolic relaxation increased P-p38MAPK, P-PKCε, P-HSP27, and P-Drp1 expression, decreased calcineurin content, and normalized calcium and membrane potential parameters measured in isolated mitochondria. These effects were significantly attenuated when ETZ was administered in the presence of SB or Chel. These data show that ETZ protects the myocardium and mitochondria against ischemia-reperfusion injury through p38MAPK- and PKCε-dependent pathways and reinforces the role of CA as a possible target in the management of acute cardiac ischemic diseases.
我们之前已经证明,抑制细胞外定向碳酸酐酶(CA)同工型可保护心肌免受缺血再灌注损伤。在这项研究中,我们的目的是通过检查高度扩散的细胞膜通透 CA 抑制剂乙氧唑胺(ETZ)的作用,评估 CA 细胞内同工型可能的进一步贡献。稳定 20 分钟后,将离体大鼠心脏分为以下几组:(1)非缺血对照组:灌注 90 分钟;(2)缺血对照组:30 分钟整体缺血和 60 分钟再灌注(R);和(3)ETZ:ETZ 在缺血开始前 10 分钟以 100 μM 的浓度给药,然后在再灌注的前 10 分钟内给药。在其他组中,ETZ 在 SB202190(SB,p38MAPK 抑制剂)或 Chelerythrine(Chel,蛋白激酶 C [PKC] 抑制剂)存在下给药。评估梗塞面积、心肌功能以及磷酸化 p38MAPK、PKCε、HSP27 和 Drp1 形式和钙调神经磷酸酶 Aβ含量的表达。在分离的线粒体中,测量 Ca 反应、Ca 保留能力和膜电位。ETZ 使梗塞面积减少 60%,改善缺血后心肌收缩和舒张松弛的恢复,增加 P-p38MAPK、P-PKCε、P-HSP27 和 P-Drp1 的表达,减少钙调神经磷酸酶含量,并使分离的线粒体中测量的 Ca 和膜电位参数正常化。当 ETZ 在 SB 或 Chel 存在下给药时,这些作用明显减弱。这些数据表明,ETZ 通过 p38MAPK 和 PKCε 依赖性途径保护心肌和线粒体免受缺血再灌注损伤,并加强 CA 作为急性心脏缺血性疾病管理的可能靶标的作用。