Research Institute for the Biology of Farm Animals, Dummerstorf, Germany.
Cytometry A. 2010 Apr;77(4):310-20. doi: 10.1002/cyto.a.20873.
N-acetylcysteine (NAC) is known as an antioxidant and used for mucus viscosity reduction. However, this drug prevents or induces cell death depending on the cell type. The response of steroidogenic luteal cells to NAC is unknown. Our data shows that NAC can behave as an antioxidant or prooxidant in dependency on the concentration and mitochondrial energization. NAC elevated the flowcytometric-measured portion of hypodiploid (dying) cells. This rise was completely abolished by aurintricarboxylic acid, an inhibitor of topoisomerase II. NAC increased the secretion of nitric oxide and cellular nitrotyrosine. An image analysis indicated that cells pretreated with NAC and loaded with DHR showed a fluorescent structure probably elicited by the oxidative product of DHR, rhodamine 123 that sequesters mitochondrially. Pretreating luteal cells with NAC or adding NAC directly to mitochondrial fractions followed by assessing the mitochondrial transmembrane potential difference (Deltapsi) by the JC-1 technique demonstrated a marked decrease in Deltapsi. A protonophore restored Deltapsi and rotenone (an inhibitor of respiratory chain complex I) inhibited mitochondrial recovering. Thus, in steroidogenic luteal cells from healthy mature corpus luteum, NAC impairs cellular survival by interfering with mitochondrial metabolism. The protonophore-induced recovering of NAC-provoked decrease in Deltapsi indicates that an ATP synthase-favored route of H(+) re-entry to the matrix is essentially switched off by NAC while other respiratory chain complexes remain intact. These data may be important for therapeutic timing of treatments with NAC. (c) 2010 International Society for Advancement of Cytometry.
N-乙酰半胱氨酸（NAC）是一种抗氧化剂，用于降低黏液的黏度。然而，这种药物的细胞毒性取决于细胞类型。关于 NAC 对甾体生成黄体细胞的作用，目前还不清楚。我们的数据显示，NAC 可以作为一种抗氧化剂或促氧化剂，取决于浓度和线粒体的能量状态。NAC 增加了流式细胞术测量的亚二倍体（凋亡）细胞的比例。这个增加被拓扑异构酶 II 抑制剂金雀异黄素完全消除。NAC 增加了一氧化氮和细胞硝基酪氨酸的分泌。图像分析表明，用 NAC 预处理并加载 DHR 的细胞显示出一种荧光结构，可能是由 DHR 的氧化产物引起的，罗丹明 123 可隔离线粒体。用 NAC 预处理黄体细胞或直接将 NAC 添加到线粒体部分，然后用 JC-1 技术评估线粒体跨膜电位差（Deltapsi），结果表明 Deltapsi 明显下降。质子载体恢复了 Deltapsi，而鱼藤酮（呼吸链复合物 I 的抑制剂）抑制了线粒体的恢复。因此，在来自健康成熟黄体的甾体生成黄体细胞中，NAC 通过干扰线粒体代谢来损害细胞存活。质子载体诱导的 NAC 引起的 Deltapsi 下降的恢复表明，NAC 基本上关闭了有利于 ATP 合酶的 H(+) 进入基质的途径，而其他呼吸链复合物保持完整。这些数据对于 NAC 治疗的治疗时机可能很重要。（c）2010 年国际细胞分析学会。
