Role of heat shock protein 90 and endothelial nitric oxide synthase during early anesthetic and ischemic preconditioning.
Role of heat shock protein 90 and endothelial nitric oxide synthase during early anesthetic and ischemic preconditioning.
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DOI：
10.1097/aln.0b013e3181942cb4
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发表时间：
2009-02
期刊：
影响因子：
8.8
通讯作者：
Kersten JR
中科院分区：
文献类型：
--
作者：
Amour J;Brzezinska AK;Weihrauch D;Billstrom AR;Zielonka J;Krolikowski JG;Bienengraeber MW;Warltier DC;Pratt PF Jr;Kersten JR
一氧化氮是心肌早期麻醉（APC）和缺血（IPC）预适应的重要组成部分。热休克蛋白90（Hsp 90）调节内皮型一氧化氮合酶（eNOS）活性。在这项研究中，我们测试的假设，热休克蛋白90-eNOS相互作用调节APC和IPC。在不存在或存在异氟烷（APC）30 min或冠状动脉闭塞（IPC）5 min预处理的情况下，测量兔冠状动脉闭塞和再灌注后的心肌梗死面积，并使用或不使用格尔德霉素或根赤霉素（两种化学上不同的Hsp 90抑制剂）或NG-硝基-L-精氨酸甲酯（一种非特异性NOS抑制剂）进行预处理。异氟醚依赖性一氧化氮的生产测定（臭氧化学发光）在人冠状动脉内皮细胞或小鼠心肌细胞，在存在或不存在热休克蛋白90抑制剂或NG-硝基-L-精氨酸甲酯。热休克蛋白90和eNOS之间的相互作用，eNOS激活进行了评估，免疫沉淀，免疫印迹，共聚焦显微镜。APC和IPC减少梗死面积（分别为50%和59%），这种作用被Hsp 90抑制剂所消除。NG-硝基-L-精氨酸甲酯阻断APC，但不阻断IPC。异氟烷增加人冠状动脉内皮细胞中一氧化氮的产生，同时增加热休克蛋白90-eNOS相互作用（免疫沉淀，免疫印迹和免疫组织化学）。用Hsp 90抑制剂预处理可阻断异氟醚依赖的一氧化氮生成，并降低Hsp 90-eNOS相互作用。异氟烷没有增加小鼠心肌细胞中一氧化氮的产生，eNOS低于检测水平。结果表明，热休克蛋白90通过蛋白质-蛋白质相互作用在介导APC和IPC中起着关键作用，并表明内皮细胞是APC过程中一氧化氮介导的信号传导的重要贡献者。
Nitric oxide is known to be essential for early anesthetic (APC) and ischemic (IPC) preconditioning of myocardium. Heat shock protein 90 (Hsp90) regulates endothelial nitric oxide synthase (eNOS) activity. In this study, we tested the hypothesis that Hsp90-eNOS interactions modulate APC and IPC. Myocardial infarct size was measured in rabbits after coronary occlusion and reperfusion in the absence or presence of preconditioning with 30 min of isoflurane (APC) or 5 min of coronary artery occlusion (IPC), and with or without pre-treatment with geldanamycin or radicicol, two chemically distinct Hsp90 inhibitors, or NG-nitro-L-arginine methylester, a non-specific NOS inhibitor. Isoflurane-dependent nitric oxide production was measured (ozone chemiluminescence) in human coronary artery endothelial cells or mouse cardiomyocytes, in the absence or presence of Hsp90 inhibitors or NG-nitro-L-arginine methylester. Interactions between Hsp90 and eNOS, and eNOS activation were assessed with immunoprecipitation, immunoblotting, and confocal microscopy. APC and IPC decreased infarct size (50% and 59%, respectively) and this action was abolished by Hsp90 inhibitors. NG-nitro-L-arginine methylester blocked APC but not IPC. Isoflurane increased nitric oxide production in human coronary artery endothelial cells, concomitantly with an increase in Hsp90-eNOS interaction (immunoprecipitation, immunoblotting, and immunohistochemistry). Pretreatment with Hsp90 inhibitors abolished isoflurane-dependent nitric oxide production and decreased Hsp90-eNOS interactions. Isoflurane did not increase nitric oxide production in mouse cardiomyocytes and eNOS was below the level of detection. The results indicate that Hsp90 plays a critical role in mediating APC and IPC through protein-protein interactions, and suggest that endothelial cells are important contributors to nitric oxide-mediated signalling during APC.