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兔脊髓缺血中的自由基:电子自旋共振光谱及其与超氧化物歧化酶活性的相关性

Free radicals in rabbit spinal cord ischemia: electron spin resonance spectroscopy and correlation with SOD activity.

作者信息

Lombardi V, Valko L, Stolc S, Valko M, Ondrejicková O, Horáková L, Placek J, Troncone A

机构信息

Department of Pathological Anatomy, Bari Oncology Center, Italy.

出版信息

Cell Mol Neurobiol. 1998 Aug;18(4):399-412. doi: 10.1023/a:1022597431593.

Abstract
  1. In nonanesthetized rabbits temporal occlusion of the abdominal aorta was used to induce oxidative stress in the lower part of the body including distal segments of the spinal cord. 2. Spinal cord samples were taken from the animals exposed to 25-min aortic occlusion (AO) or to occlusion followed by 1- or 2-hr reperfusion (AO/R1 or AO/R2, respectively) or from sham-operated animals (C). The presence of free radicals (FR) in the spinal cord samples frozen in liquid N2 was assessed by ESR spectroscopy without spin trapping. Moreover, superoxide dismutase (SOD) activity and conjugated diene (CD) levels were measured in the samples. 3. In the AO group FR were detected in the spinal cord regions close to the occlusion (lower thoracic and distal segments) along with a decrease in SOD activity. The calculated g value (g = 2.0291) indicated that the paramagnetic signal recorded might be attributed to superoxide radicals. FR were absent in the AO/R1 group. Concurrently, the SOD activity revealed a significant tendency to return to the control level. FR appeared again in the AO/R2 group, mostly in the upper and middle lumbar regions, along with a decrease in SOD activity. No sample from the C group revealed FR. A significant increase in CD levels was observed in the thoracolumbar region only in the AO/R2 group. The temporary absence of FR in the AO/R1 group suggests activation of defense antioxidant mechanisms (e.g., specific enzymatic systems such as SOD), which might have been exhausted later. 4. Changes in SOD activity similar to those observed in the thoracolumbar region, though less noticeable, occurred in the obviously noncompromised tissue (upper cervical region). This points to a kind of generalized response of the animal to aortic occlusion. 5. Direct ESR spectroscopy revealed the presence of FR as well as their time course in the spinal cord during the early phase of ischemia/reperfusion injury and the inverse relationship between FR and SOD activity.
摘要
  1. 在未麻醉的兔子中,通过对腹主动脉进行颞部闭塞来诱导身体下部包括脊髓远端节段的氧化应激。2. 从暴露于25分钟主动脉闭塞(AO)或闭塞后再灌注1小时或2小时(分别为AO/R1或AO/R2)的动物或假手术动物(C)中采集脊髓样本。通过无自旋捕获的电子自旋共振光谱法评估液氮冷冻的脊髓样本中自由基(FR)的存在。此外,还测量了样本中的超氧化物歧化酶(SOD)活性和共轭二烯(CD)水平。3. 在AO组中,在靠近闭塞部位的脊髓区域(下胸段和远端节段)检测到FR,同时SOD活性降低。计算出的g值(g = 2.0291)表明记录到的顺磁信号可能归因于超氧自由基。AO/R1组中未检测到FR。同时,SOD活性显示出显著的恢复到对照水平的趋势。FR在AO/R2组中再次出现,主要在上腰部和中腰部区域,同时SOD活性降低。C组样本中未检测到FR。仅在AO/R2组的胸腰段区域观察到CD水平显著升高。AO/R1组中FR的暂时缺失表明防御性抗氧化机制(例如特定的酶系统如SOD)被激活,而这些机制可能随后会耗尽。4. 在明显未受损的组织(上颈段区域)中发生了与胸腰段区域类似的SOD活性变化,尽管不太明显。这表明动物对主动脉闭塞有一种全身性的反应。5. 直接电子自旋共振光谱法揭示了缺血/再灌注损伤早期脊髓中FR的存在及其时间进程,以及FR与SOD活性之间的反比关系。

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