Sevoflurane postconditioning protects isolated rat hearts against ischemia-reperfusion injury
Sevoflurane postconditioning protects isolated rat hearts against ischemia-reperfusion injury
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DOI：
10.3760/cma.j.issn.0366-6999.2010.10.018
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发表时间：
2010-05-20
影响因子：
6.1
通讯作者：
Li Li-huan
中科院分区：
文献类型：
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作者：
Yao Yun-tai;Fang Neng-xin;Li Li-huan
背景研究表明，在再灌注早期或“麻醉后处理”时使用麻醉药可以保护缺血后心脏免受心肌缺血再灌注损伤(MIRO)。然而，负责这种保护的机制并没有得到很好的阐明。通过选择性抑制PI3K、ERK 1/2和线粒体通透性转换孔(MPTP)，分别用LY294002(LY)、PD98059(PD)、5-Hd(5-HD)阻断PI3K(LY)、PD98059(PD)、5-Hd(5-HD)，观察七氟烷后处理(SpostC)对体外培养大鼠心脏的保护作用，以及PI3K、ERK 1/2和线粒体通透性转换孔(MPTP)的作用。方法将大鼠离体心随机分为12组(n=15)：时间对照组(持续灌流)、ISCH组(缺血30min后再灌流60min)、SpostC组(缺血30min后再灌流前15分钟给予3%七氟醚后处理)、ISCH+LY、ISCH+PD、ISCH+ATR、ISCH+5-HD、ISCH+DMSO组(LY、PD、ATR、5-HD和DMSO)、SpostC+LY、SpostC+PD、SpostC+ATR和SpostC+5-HD组(LY、PD、DMSO)。ATR和5-HD分别与3%七氟醚合用)。比较组内和组间的血流动力学。实验结束时用氯化三苯基四氮唑(TTC)染色测定心肌梗死面积。比较TC组、ISCH组和SpostC组心肌组织乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)和心肌肌钙蛋白1(CTnI)的变化。为探讨MPTP参与SpostC的危险性，比较了不同实验组(TO、ISCH、ISCH+LY、ISCH+PD、ISCH+DMSO、SpostC、SpostC+LY、SpostC+PD)心肌组织中MPTP开放的标志NAD+含量。为进一步探讨抗凋亡机制是否参与SpostC诱导的心肌保护及其与线粒体的关系，对部分实验组(TO组、ISCH组、ISCH+5-HD组、ISCH+ATR组、ISch+DMSO组、SpostC组、SpostC+5-HD组、SpostC+ATR组)进行TUNEL染色。结果与缺血30min的未保护心脏相比，3%七氟醚在再灌注早期15min可显著促进心肌功能恢复，缩小心肌梗死范围，减少LDH、OK-MB和cTnI的释放，并减少心肌细胞凋亡(P
Background Studies suggested that anesthetics administered upon the early reperfusion or "anesthetic postconditioning" could protect post-ischemic hearts against myocardial ischemia reperfusion injury (MIRO. However, the mechanism responsible for such protection was not well-elucidated. We investigated the cardioprotection induced by sevoflurane postconditioning (SpostC) in rat hearts in vitro, and the respective role of phosphatidylinositol-3-kinase (PI3K), extracellular signal-regulated kinase 1 and 2 (ERK 1/2), mitochondrial K-ATP channels (mitoK(ATP)) and mitochondrial permeability transition pore (mPTP), by selectively inhibiting PI3K, ERK 1/2, mitoK(ATP), with LY294002 (LY), PD98059 (PD), 5-hydroxydecanoate (5-HD) and by directly opening of mPTP with atractyloside (ATR), respectively.Methods Isolated rat hearts were randomly assigned to one of the 12 groups (n=15): Time control (continuous perfusion), ISCH (30 minutes of ischemia followed by 60 minutes of reperfusion alone), SpostC (3% sevoflurane postconditioning was administered during the first 15 minutes of reperfusion after 30 minutes of ischemia), ISCH+LY, ISCH+PD, ISCH+ATR, ISCH+5-HD and ISCH+ dimethyl sulfoxide (DMSO) groups (LY, PD, ATR, 5-HD and DMSO (the vehicle) was administered respectively during the first 15 minutes of reperfusion following test ischemia), SpostC+LY, SpostC+PD, SpostC+ATR and SpostC+5-HD groups (LY, PD, ATR and 5-HD was coadministered with 3% sevoflurane, respectively). Hemodynamics was compared within and between groups. Infarction size was determined at the end of experiments using triphenyltetrazolium chloride (TTC) staining. Lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB) and cardiac troponin 1 (cTnI) released from necrotic myocardium, were compared among TC, ISCH and SpostC groups. To investigate the relationships between RISK and mPTP implicated in SpostC, NAD+ content in myocardium, a marker of mPTP opening, was compared among some experimental groups (TO, ISCH, ISCH+LY, ISCH+PD, ISCH+DMSO, SpostC, SpostC+LY, SpostC+PD). To further investigate whether the anti-apoptotic mechanism is implicated in SpostC-induced cardioprotection and its association with mitochondria, TUNEL staining was performed in some experimental groups (TO, ISCH, ISCH+5-HD, ISCH+ATR, ISCH+DMSO, SpostC, SpostC+5-HD, SpostC+ATR).Results When compared with unprotected hearts subjected to 30 minutes of ischemia, exposure to 3% sevoflurane for 15 minutes during early reperfusion significantly improved functional recovery, decreased myocardial infarct size, decreased LDH, OK-MB and cTnI release, and decreased cardiomyocyte apoptosis (P