Antioxidant defences and homeostasis of reactive oxygen species in different human mitochondrial DNA-depleted cell lines.

Lodovica Vergani, Maura Floreani, Aaron Russell, Mara Ceccon, Eleonora Napoli, Anna Cabrelle, Lucia Valente, Federica Bragantini, Bertrand Leger, Federica Dabbeni-Sala
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引用次数: 56

Abstract

Three pairs of parental (rho+) and established mitochondrial DNA depleted (rho0) cells, derived from bone, lung and muscle were used to verify the influence of the nuclear background and the lack of efficient mitochondrial respiratory chain on antioxidant defences and homeostasis of intracellular reactive oxygen species (ROS). Mitochondrial DNA depletion significantly lowered glutathione reductase activity, glutathione (GSH) content, and consistently altered the GSH2 : oxidized glutathione ratio in all of the rho0 cell lines, albeit to differing extents, indicating the most oxidized redox state in bone rho0 cells. Activity, as well as gene expression and protein content, of superoxide dismutase showed a decrease in bone and muscle rho0 cell lines but not in lung rho0 cells. GSH peroxidase activity was four times higher in all three rho0 cell lines in comparison to the parental rho+, suggesting that this may be a necessary adaptation for survival without a functional respiratory chain. Taken together, these data suggest that the lack of respiratory chain prompts the cells to reduce their need for antioxidant defences in a tissue-specific manner, exposing them to a major risk of oxidative injury. In fact bone-derived rho0 cells displayed the highest steady-state level of intracellular ROS (measured directly by 2',7'-dichlorofluorescin, or indirectly by aconitase activity) compared to all the other rho+ and rho0 cells, both in the presence or absence of glucose. Analysis of mitochondrial and cytosolic/iron regulatory protein-1 aconitase indicated that most ROS of bone rho0 cells originate from sources other than mitochondria.

不同人类线粒体dna缺失细胞系中活性氧的抗氧化防御和体内平衡。
本研究利用来自骨、肺和肌肉的三对亲本(rho+)和已建立的线粒体DNA缺失(rho0)细胞来验证核背景和缺乏有效线粒体呼吸链对细胞内活性氧(ROS)的抗氧化防御和稳态的影响。线粒体DNA缺失显著降低了所有rho0细胞系中谷胱甘肽还原酶活性和谷胱甘肽(GSH)含量,并持续改变了GSH2:氧化谷胱甘肽的比例,尽管程度不同,表明骨rho0细胞中氧化氧化还原状态最严重。骨和肌rho0细胞超氧化物歧化酶活性、基因表达和蛋白含量均下降,肺rho0细胞无明显变化。GSH过氧化物酶活性在所有3个rho0细胞系中比亲本rho+细胞系高4倍,这表明这可能是在没有功能呼吸链的情况下生存的必要适应。综上所述,这些数据表明,缺乏呼吸链会促使细胞以一种组织特异性的方式减少对抗氧化防御的需求,从而使它们面临氧化损伤的主要风险。事实上,与所有其他rho+和rho0细胞相比,骨源性rho0细胞显示出最高的稳态细胞内ROS水平(直接通过2',7'-二氯荧光素测量,或间接通过乌头酶活性测量),无论是否存在葡萄糖。线粒体和细胞质/铁调节蛋白-1乌头酸酶的分析表明,骨rho0细胞的大多数ROS来源于线粒体以外的来源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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