Pieri C, Moroni F, Recchioni R
Cytology Center, Gerontol. Res. Dept. of I.N.R.C.A., Ancona, Italy.
Free Radic Biol Med. 1993 Dec;15(6):661-5. doi: 10.1016/0891-5849(93)90170-y.
This study was designed to evaluate the time-dependent changes of mitochondrial membrane potential and mass during Con-A-induced proliferation of splenic lymphocytes from rat fed a normal or a vitamin E deficient diet. Rhodamine 123 and Nonyl Acridine Orange were used as specific probes to monitor the membrane potential and mass of mitochondria, respectively, by means of flow cytometry. The results demonstrate that the increase of Rh-123 and NAO uptake observed in cells from normally fed rats was prevented by vitamin E deficiency, at any time considered. After 72 h from Con A stimulation, 62% of cells from controls, as against 16% of cells from vitamin E deficient rats, showed hyperpolarized mitochondria. At the same time, in this last group, 60% of cells had depolarized organelles. The same pattern was observed considering the changes of mitochondrial mass, measured using NAO as a probe. These data support that mitogenic stimulation induced an increase of the respiratory activity of mitochondria with subsequent production of superoxide radicals. This resulted in depolarization and loss of mass of the organelles if the intracellular level of vitamin E is not adequate.
本研究旨在评估正常饮食或维生素E缺乏饮食的大鼠脾脏淋巴细胞在刀豆蛋白A诱导增殖过程中，线粒体膜电位和质量随时间的变化。分别使用罗丹明123和壬基吖啶橙作为特异性探针，通过流式细胞术监测线粒体的膜电位和质量。结果表明，在任何时间点，维生素E缺乏均会阻止正常饮食大鼠细胞中观察到的罗丹明123摄取增加和壬基吖啶橙摄取增加。刀豆蛋白A刺激72小时后，对照组62%的细胞线粒体超极化，而维生素E缺乏大鼠组仅16%的细胞如此。同时，在维生素E缺乏大鼠组中，60%的细胞线粒体去极化。以壬基吖啶橙为探针测量线粒体质量变化时，也观察到相同模式。这些数据支持有丝分裂刺激会诱导线粒体呼吸活性增加，随后产生超氧自由基。如果细胞内维生素E水平不足，这会导致线粒体去极化和质量损失。
