MYOCARDIAL ISCHEMIA AND REPERFUSION - DIRECT EVIDENCE FOR FREE-RADICAL GENERATION BY ELECTRON-SPIN RESONANCE SPECTROSCOPY
MYOCARDIAL ISCHEMIA AND REPERFUSION - DIRECT EVIDENCE FOR FREE-RADICAL GENERATION BY ELECTRON-SPIN RESONANCE SPECTROSCOPY
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DOI：
10.1073/pnas.85.8.2786
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发表时间：
1988-04-01
影响因子：
11.1
通讯作者：
KALYANARAMAN, B
中科院分区：
文献类型：
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作者：
BAKER, JE;FELIX, CC;KALYANARAMAN, B
电子自旋共振光谱学最近已被其他人用于直接检测离体灌注心脏中的自由基种类。在这项研究中，光谱学之前的样品处理涉及粉碎组织，这可能会人为地产生自由基物质。我们评估了在离体灌注心脏的影响，组织粉碎的光谱检测到的自由基物种的身份，然后，使用处理技术不太可能诱导文物，是否心肌缺血和再灌注产生自由基物种。将大鼠和兔心脏（n = 8）有氧灌注10分钟并冷冻夹持至-196 ℃。C.冷冻组织在-196 ℃下处理。C用于通过粉碎与切碎进行光谱分析。粉末组织的光谱由三种组分组成：半醌（g = 2.004），脂质过氧自由基（g. dblvert）。= 2.04和g.perp.）= 2.006）和可能是脂质自由基的碳中心自由基（giso = 2.002和AzzH = 2.006）。50 G）。切碎的组织由单一组分，半醌（g = 2.004）。大鼠心脏（每组n = 8）也经历了10分钟的全球无流量常温缺血，然后5-60秒的有氧或无氧再灌注，冷冻组织切碎，然后进行光谱学。缺血组织的光谱由铁硫中心和半醌组成。有氧再灌注导致与对照相似的频谱，但具有增加的幅度，产生与缺血组织相同的频谱。我们得出结论，粉碎冷冻心肌组织人为产生自由基物种。使用非粉碎技术的组织处理，我们发现，心肌缺血和再灌注产生自由基的物种，但分子氧是必要的自由基生产过程中流出的爆发。
Electron spin resonance spectroscopy has recently been used by others to detect directly radical species in isolated perfused hearts. Sample processing prior to spectroscopy in this study involved pulverization of tissue, which can artifactually generate radical species. We assessed in isolated perfused hearts the influence of tissue pulverization on the identity of radical species detected by spectroscopy and then, using a processing technique less likely to induce artifacts, whether myocardial ischemia and reperfusion generate radical species. Rat and rabbit hearts (n = 8) were perfused aerobically for 10 min and freeze-clamped to -196.degree. C. Frozen tissue was processed at -196.degree. C for spectroscopic analysis by pulverization vs. chopping. Spectra of pulverized tissue consisted of three components: a semiquinone (g = 2.004), a lipid peroxy radical (g.dblvert. = 2.04 and g.perp.) = 2.006), and a carbon-centered radical that is possibly a lipid radical (giso = 2.002 and AzzH .apprxeq. 50 G). Chopped tissue consisted of a single component, a semiquinone (g = 2.004). Rat hearts (n = 8 per group) also underwent 10-min global no-flow normothermic ischemia followed by 5-60 sec of either aerobic or anaerobic reperfusion, with frozen tissue chopped prior to spectroscopy. Spectra of ischemic tissue consisted of an iron-sulfur center and a semiquinone. Aerobic reperfusion resulted in a spectrum similar to the control but with increased amplitude that yielded a spectrum identical to that of ischemic tissue. We conclude that pulverization of frozen myocardial tissue artifactually generates radical species. Using a nonpulverization technique for tissue processing, we found that myocardial ischemia and reperfusion produce radical species but that molecular oxygen is necessary for the burst of radical production during outflow.