人体骨骼肌线粒体体外氧电极研究。重新评估。

Oxygen electrode studies with human skeletal muscle mitochondria in vitro. A re-appraisal.

作者信息

Byrne E, Trounce I

出版信息

J Neurol Sci. 1985 Jul;69(3):319-33. doi: 10.1016/0022-510x(85)90143-1.

Abstract

Polarographic measurement of respiration rates in skeletal muscle mitochondria is a valuable research tool in human disease but the data on normal muscle are sparse and conflicting. In this study, mitochondria were isolated from 18 normal muscle biopsies and state 3 respiration rates measured with glutamate/malate, pyruvate/malate and succinate as substrates. In 13 experiments, reproducible respiration rates with adequate respiratory control ratios (RCR) and ADP/O ratios were obtained. Mean state 3 respiration rates of 280 ng-atoms 0 min-1 mg mitochondrial protein-1 (30 degrees C) with glutamate, 214 ng-atoms 0 min-1 mg mitochondrial protein-1 (30 degrees C) with pyruvate and 353 ng-atoms 0 min-1 mg mitochondrial protein-1 (30 degrees C) with succinate as substrates were higher than in most earlier reports. Selective uncoupling with alpha-glycerophosphate and data suggesting a partial site I deficiency were found in some experiments where RCR and ADP/O ratio measurements indicated a good preparation. Slow rates with all substrates with loose coupling were found in 3 studies where the mitochondria were clearly damaged during extraction. Guidelines for the evaluation of polarographic data in reports in the literature are suggested.

摘要

极谱法测量骨骼肌线粒体的呼吸速率是研究人类疾病的一项重要研究工具，但关于正常肌肉的相关数据稀少且相互矛盾。在本研究中，从18份正常肌肉活检样本中分离出线粒体，并以谷氨酸/苹果酸、丙酮酸/苹果酸和琥珀酸作为底物测量状态3呼吸速率。在13项实验中，获得了具有足够呼吸控制率（RCR）和ADP/O比率的可重复呼吸速率。以谷氨酸为底物时，状态3平均呼吸速率为280纳克原子氧·分钟⁻¹·毫克线粒体蛋白⁻¹（30℃），以丙酮酸为底物时为214纳克原子氧·分钟⁻¹·毫克线粒体蛋白⁻¹（30℃），以琥珀酸为底物时为353纳克原子氧·分钟⁻¹·毫克线粒体蛋白⁻¹（30℃），均高于大多数早期报告中的数据。在一些RCR和ADP/O比率测量显示制备良好的实验中，发现了与α-甘油磷酸的选择性解偶联以及提示部分位点I缺陷的数据。在3项研究中，发现所有底物的速率缓慢且偶联松散，这些研究中线粒体在提取过程中明显受损。文中还提出了文献报告中极谱数据评估的指导原则。