Urban Nadine, Tsitsipatis Dimitrios, Hausig Franziska, Kreuzer Katrin, Erler Katrin, Stein Vanessa, Ristow Michael, Steinbrenner Holger, Klotz Lars-Oliver
Institute of Nutrition, Department of Nutrigenomics, Friedrich-Schiller-Universität Jena, D-07743 Jena, Germany.
ETH Zurich (Swiss Federal Institute of Technology), Schorenstrasse 16, Building SLA C7, CH-8603 Schwerzenbach-Zurich, Switzerland.
Redox Biol. 2017 Apr;11:502-515. doi: 10.1016/j.redox.2016.12.003. Epub 2016 Dec 6.
The redox environment in cells and organisms is set by low-molecular mass and protein-bound thiols, with glutathione (GSH) representing a major intracellular redox buffer. Subtle thiol oxidation elicits signal transduction processes and adaptive responses to cope with stressors, whereas highly oxidizing conditions may provoke cell death. We here tested how thiol depletion affects life span, stress resistance and stress signaling in the model organism Caenorhabditis elegans. Diethyl maleate (DEM), an α,β-unsaturated carbonyl compound that conjugates to GSH and other thiols, decreased C. elegans life span at a concentration of 1mM. In contrast, low and moderate doses of DEM (10-100µM) increased mean and maximum life span and improved resistance against oxidative stress. DEM-induced life span extension was not detectable in worms deficient in either the FoxO orthologue, DAF-16, or the Nrf2 orthologue, SKN-1, pointing to a collaborative role of the two transcription factors in life span extension induced by thiol depletion. Cytoprotective target genes of DAF-16 and SKN-1 were upregulated after at least 3 days of exposure to 100µM DEM, but not 1mM DEM, whereas only 1mM DEM caused upregulation of egl-1, a gene controlled by a p53-orthologue, CEP-1. In order to test whether depletion of GSH may elicit effects similar to DEM, we suppressed GSH biosynthesis in worms by attenuating γ-glutamylcysteine synthetase (gcs-1) expression through RNAi. The decline in GSH levels elicited by gcs-1 knockdown starting at young adult stage did not impair viability, but increased both stress resistance and life expectancy of the worms. In contrast, gcs-1 knockdown commencing right after hatching impaired nematode stress resistance and rendered young adult worms prone to vulval ruptures during egg-laying. Thus, modest decrease in GSH levels in young adult worms may promote stress resistance and life span, whereas depletion of GSH is detrimental to freshly hatched and developing worms.
细胞和生物体中的氧化还原环境由低分子量和与蛋白质结合的硫醇设定,谷胱甘肽(GSH)是主要的细胞内氧化还原缓冲剂。细微的硫醇氧化引发信号转导过程和对压力源的适应性反应,而高度氧化的条件可能引发细胞死亡。我们在此测试了硫醇耗竭如何影响模式生物秀丽隐杆线虫的寿命、抗逆性和应激信号传导。马来酸二乙酯(DEM)是一种与GSH和其他硫醇结合的α,β-不饱和羰基化合物,在1mM浓度下会缩短秀丽隐杆线虫的寿命。相比之下,低剂量和中等剂量的DEM(10 - 100µM)可延长平均寿命和最大寿命,并提高对氧化应激的抵抗力。在缺乏FoxO同源物DAF - 16或Nrf2同源物SKN - 1的线虫中,未检测到DEM诱导的寿命延长,这表明这两种转录因子在硫醇耗竭诱导的寿命延长中具有协同作用。在暴露于100µM DEM至少3天后,但不是1mM DEM,DAF - 16和SKN - 1的细胞保护靶基因上调,而只有1mM DEM导致egl - 1上调,egl - 1是一个由p53同源物CEP - 1控制的基因。为了测试GSH耗竭是否可能引发与DEM类似的效应,我们通过RNAi减弱γ-谷氨酰半胱氨酸合成酶(gcs - 1)的表达来抑制线虫中的GSH生物合成。从成虫早期开始,gcs - 1敲低引起的GSH水平下降并未损害线虫的活力,但增加了线虫的抗逆性和预期寿命。相比之下,孵化后立即开始敲低gcs - 1会损害线虫的抗逆性,并使成虫在产卵期间容易出现外阴破裂。因此,成虫中GSH水平的适度降低可能促进抗逆性和寿命,而GSH的耗竭对刚孵化和发育中的线虫是有害的。