Yurkow E J, McKenzie M A
Department of Pharmacology and Toxicology, Rutgers University, Piscataway, New Jersey 08854.
Cytometry. 1993;14(3):287-93. doi: 10.1002/cyto.990140309.
Peroxide production in cultures of Saccharomyces cerevisiae was measured using the H2O2-sensitive fluorescent probe 2',7'-dichlorofluorescein diacetate (DCFH-DA) and flow cytometry. Aeration of cultures of S. cerevisiae exposed to a period of hypoxia was found to induce elevated levels of peroxide that were 100-fold higher than the levels observed in cultures maintained under exclusively aerated or hypoxic conditions. Simultaneous viability analysis, using the fluorescent DNA-intercalating dye propidium iodide, indicated that the increase in peroxide generation preceded cell damage and death. Various agents were found to influence the effect of peroxides on cell viability. The addition of ethanol to hypoxic stationary cultures dramatically increased the rate of cell death without further increasing the amount of peroxide produced, while glucose inhibited peroxide production and decreased the rate of cell death. Surprisingly, elevated peroxide levels of hypoxic/reaerated cultures were maintained upon addition of KH2PO4, although the cells remained viable for extended periods of time when compared to control and other test cultures. Similarities between our observations and those of other investigators using anoxic/reperfused organs suggest that hypoxic/reaerated yeast cultures may be a useful model system to study ischemia-dependent tissue destruction of mammals.
使用对过氧化氢敏感的荧光探针2',7'-二氯荧光素二乙酸酯（DCFH-DA）和流式细胞术来测量酿酒酵母培养物中过氧化氢的产生。发现暴露于一段缺氧时期的酿酒酵母培养物通气会诱导过氧化氢水平升高，该水平比在完全通气或缺氧条件下培养的培养物中观察到的水平高100倍。使用荧光DNA嵌入染料碘化丙啶进行的同步活力分析表明，过氧化氢生成的增加先于细胞损伤和死亡。发现各种试剂会影响过氧化物对细胞活力的作用。向缺氧的静止培养物中添加乙醇会显著增加细胞死亡率，而不会进一步增加产生的过氧化物量，而葡萄糖会抑制过氧化物产生并降低细胞死亡率。令人惊讶的是，添加KH2PO4后，缺氧/再通气培养物中过氧化物水平仍保持升高，尽管与对照和其他测试培养物相比，细胞在较长时间内仍保持活力。我们的观察结果与其他使用缺氧/再灌注器官的研究人员的观察结果之间的相似性表明，缺氧/再通气酵母培养物可能是研究哺乳动物缺血依赖性组织破坏的有用模型系统。
