Evaluation of mitochondrial respiratory function in small biopsies of liver
Evaluation of mitochondrial respiratory function in small biopsies of liver
复制标题
DOI：
10.1006/abio.2002.5658
复制
发表时间：
2002-06-15
影响因子：
2.9
通讯作者：
Gnaiger, E
中科院分区：
文献类型：
--
作者：
Kuznetsov, AV;Strobl, D;Gnaiger, E
在透化的猪肝活组织检查中研究线粒体呼吸功能。在2-7 mg的组织样品中机械透化细胞膜，用于在高分辨率呼吸测定法中应用标准化底物/抑制剂滴定方案。特定的呼吸测试表明完整的质膜透性和访问的基板完整的线粒体。高呼吸腺苷酸控制率和细胞色素c的保护在组织制备中是可比的，甚至更好地比在分离的线粒体。柠檬酸合酶和细胞色素c氧化酶活性保持在85%的控制后，长达98小时的存储肝组织在0 degreesC的组氨酸-色氨酸-酮戊二酸溶液。然而，多个线粒体缺陷，在48小时冷藏后，由呼吸能力的下降，这是降低到更大的程度与复合物I基板相比，呼吸与复合物II或IV的基板，在细胞色素C的情况下测量。长时间缺血后，腺苷酸控制率显著降低，细胞色素c的刺激作用检测到细胞色素c耗竭。高分辨率呼吸测定法允许在几毫克透化肝组织中评估线粒体功能，而无需分离线粒体。这为分析人肝活检中线粒体功能提供了基础。(C)2002 Elsevier Science（美国）。
Mitochondrial respiratory function was studied in permeabilized pig liver biopsies. The cell membrane was permeabilized mechanically in tissue samples of 2-7 mg, for application of a standardized substrate/inhibitor titration protocol in high-resolution respirometry. Specific respirometric tests demonstrated complete plasma membrane permeabilization and accessibility of substrates to intact mitochondria. High respiratory adenylate control ratios and cytochrome c conservation in the tissue preparation were comparable or even better than in isolated mitochondria. Citrate synthase and cytochrome c oxidase activities remained at 85% of controls after up to 98 h storage of liver tissue at 0degreesC in histidine-tryptophan-ketoglutarate solution. Multiple mitochondrial defects, however, were indicated after 48 h cold storage by the decline in respiratory capacity, which was lowered to a larger extent with complex I substrates compared to respiration with substrates for complex II or IV, measured in the absence of cytochrome c. After prolonged ischemia, the adenylate control ratio was significantly reduced, and cytochrome c depletion was detected by the stimulatory effect of cytochrome c. High-resolution respirometry allows the assessment of mitochondrial function in a few milligrams of permeabilized liver tissue, without isolation of mitochondria. This provides a basis for the analysis of mitochondrial function in human liver biopsies. (C) 2002 Elsevier Science (USA).