Milrinone-Induced Pharmacological Preconditioning in Cardioprotection: Hints for a Role of Mitochondrial Mechanisms
Milrinone-Induced Pharmacological Preconditioning in Cardioprotection: Hints for a Role of Mitochondrial Mechanisms
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DOI：
10.3390/jcm8040507
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发表时间：
2019-04-01
影响因子：
3.9
通讯作者：
Bunte, Sebastian
中科院分区：
文献类型：
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作者：
Raupach, Annika;Reinle, Julia;Bunte, Sebastian
线粒体钙敏感性钾通道（mBK（Ca））的激活在预处理诱导的心肌保护中起着关键作用。对于米力农（Mil）诱导的预处理，参与的mBK（Ca）-通道和进一步的线粒体信号是未知的。我们推测：（1）Mil诱导的预处理是浓度依赖性的;（2）可能参与了mBK（Ca）-通道的激活、活性氧（ROS）的释放和线粒体通透性转换孔（mPTP）。用Krebs-Henseleit缓冲液灌注雄性Wistar大鼠离体心脏，缺血33 min，再灌注60 min。为了测定Mil的浓度依赖性作用，在缺血前用不同浓度的Mil（0.3-10 μ M）灌注心脏10分钟。在第二组实验中，除了对照组之外，用最低保护浓度的1 μ M Mil单独或与mBK（Ca）-通道阻断剂paxilline组合（Pax + Mil）或单独的paxilline（Pax）预处理心脏。在另外的组中，Mil分别与和不与ROS清除剂N-2-巯基丙酰甘氨酸（MPG + Mil，MPG）或mPTP抑制剂环孢菌素A（MPG + Mil + CsA，CsA + Mil）一起施用。通过氯化三苯基四氮唑（TTC）染色测定细胞大小。最低和最大的心脏保护浓度是1 μ M Mil（Mil 1：32 +/- 6%; p < 0.05 vs. Con：63 +/- 8%和Mil 0.3：49 +/- 6%）。Pax和MPG阻断了Mil的梗死面积减小（Pax + Mil：53 +/-6%，MPG + Mil：59 +/-7%; p < 0.05对比Mil：34 +/-6%），而当单独施用时对梗死面积没有影响（Pax：53 +/-7%，MPG：58 +/-5%; ns对比Con）。CsA的联合给药完全恢复了Mil的MPG抑制的心脏保护作用（MPG + Mil + CsA：35 +/-7%，p < 0.05 vs.MPG + Mil）。米力农浓度依赖性诱导预适应。钙通道的激活、ROS的释放和mPTP的抑制介导了钙通道的保护作用。
The activation of mitochondrial calcium-sensitive potassium (mBK(Ca)) channels is crucially involved in cardioprotection induced by preconditioning. For milrinone (Mil)-induced preconditioning, the involvement of mBK(Ca)-channels and further mitochondrial signaling is unknown. We hypothesize that (1) Mil-induced preconditioning is concentration-dependent and (2) that the activation of mBK(Ca)-channels, release of reactive oxygen species (ROS), and the mitochondrial permeability transition pore (mPTP) could be involved. Isolated hearts of male Wistar rats were perfused with Krebs-Henseleit buffer and underwent 33 min of ischemia followed by 60 min of reperfusion. For determination of a concentration-dependent effect of Mil, hearts were perfused with different concentrations of Mil (0.3-10 mu M) over 10 min before ischemia. In a second set of experiments, in addition to controls, hearts were pretreated with the lowest protective concentration of 1 mu M Mil either alone or combined with the mBK(Ca)-channel blocker paxilline (Pax + Mil), or paxilline alone (Pax). In additional groups, Mil was administered with and without the ROS scavenger N-2-mercaptopropionylglycine (MPG + Mil, MPG) or the mPTP inhibitor cyclosporine A (MPG + Mil + CsA, CsA + Mil), respectively. Infarct sizes were determined by triphenyltetrazolium chloride (TTC) staining. The lowest and most cardioprotective concentration was 1 mu M Mil (Mil 1: 32 +/- 6%; p < 0.05 vs. Con: 63 +/- 8% and Mil 0.3: 49 +/- 6%). Pax and MPG blocked the infarct size reduction of Mil (Pax + Mil: 53 +/- 6%, MPG + Mil: 59 +/- 7%; p < 0.05 vs. Mil: 34 +/- 6%) without having an effect on infarct size when administered alone (Pax: 53 +/- 7%, MPG: 58 +/- 5%; ns vs. Con). The combined administration of CsA completely restored the MPG-inhibited cardioprotection of Mil (MPG + Mil + CsA: 35 +/- 7%, p < 0.05 vs. MPG + Mil). Milrinone concentration-dependently induces preconditioning. Cardioprotection is mediated by the activation of mBK(Ca)-channels, release of ROS and mPTP inhibition.