Quantification of in vivo oxidative damage in Caenorhabditis elegans during aging by endogenous F3-isoprostane measurement
Quantification of in vivo oxidative damage in Caenorhabditis elegans during aging by endogenous F3-isoprostane measurement
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DOI：
10.1111/acel.12043
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发表时间：
2013-04-01
期刊：
影响因子：
7.8
通讯作者：
Brenkman, Arjan B.
中科院分区：
文献类型：
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作者：
Labuschagne, Christiaan F.;Stigter, Edwin C. A.;Brenkman, Arjan B.
氧化损伤被认为是病理和衰老发展的主要原因。然而，氧化损伤的定量在方法上是困难的。在这里，我们提出了一个强大的液相色谱串联质谱法（LC-MS/MS）的方法，准确，灵敏，线性的内源性氧化损伤的线虫体内定量，基于F3-异前列烷。F3-异前列腺素是由活性氧（ROS）引起的脂质过氧化反应引起的氧化损伤的类前列腺素标志物。在整个动物和多个细胞隔室（包括线粒体和过氧化物酶体）中定量氧化损伤。线粒体电子传递蛋白mev-1和mv-1的突变体显示氧化损伤水平增加。此外，超氧化物歧化酶（SOD）和过氧化氢酶（CTL）突变体的分析发现，氧化损伤水平不能推断从表型的抗促氧化剂单独和揭示高氧化损伤的化学感觉神经元的一个小组。老化线虫的纵向分析显示，氧化损伤增加，特别是与postreproductive年龄。值得注意的是，抗应激和长寿的daf-2胰岛素/IGF-1受体突变体的衰老涉及不同的daf-16依赖性氧化损伤阶段，包括在年轻成年时的时间增加。这些观察结果与对ROS的激素反应一致。
Oxidative damage is thought to be a major cause in development of pathologies and aging. However, quantification of oxidative damage is methodologically difficult. Here, we present a robust liquid chromatographytandem mass spectrometry (LC-MS/MS) approach for accurate, sensitive, and linear in vivo quantification of endogenous oxidative damage in the nematode Caenorhabditis elegans, based on F3-isoprostanes. F3-isoprostanes are prostaglandin-like markers of oxidative damage derived from lipid peroxidation by Reactive Oxygen Species (ROS). Oxidative damage was quantified in whole animals and in multiple cellular compartments, including mitochondria and peroxisomes. Mutants of the mitochondrial electron transport proteins mev-1 and clk-1 showed increased oxidative damage levels. Furthermore, analysis of Superoxide Dismutase (sod) and Catalase (ctl) mutants uncovered that oxidative damage levels cannot be inferred from the phenotype of resistance to pro-oxidants alone and revealed high oxidative damage in a small group of chemosensory neurons. Longitudinal analysis of aging nematodes revealed that oxidative damage increased specifically with postreproductive age. Remarkably, aging of the stress-resistant and long-lived daf-2 insulin/IGF-1 receptor mutant involved distinct daf-16-dependent phases of oxidative damage including a temporal increase at young adulthood. These observations are consistent with a hormetic response to ROS.