Han Chul, Kim Mi-Jung, Ding Dalian, Park Hyo-Jin, White Karessa, Walker Logan, Gu Tongjun, Tanokura Masaru, Yamasoba Tatsuya, Linser Paul, Salvi Richard, Someya Shinichi
Department of Aging and Geriatric Research, University of Florida, Gainesville, FL, United States of America.
Center for Hearing and Deafness, State University of New York at Buffalo, NY, United States of America.
PLoS One. 2017 Jul 7;12(7):e0180817. doi: 10.1371/journal.pone.0180817. eCollection 2017.
Glutathione reductase (GSR), a key member of the glutathione antioxidant defense system, converts oxidized glutathione (GSSG) to reduced glutathione (GSH) and maintains the intracellular glutathione redox state to protect the cells from oxidative damage. Previous reports have shown that Gsr deficiency results in defects in host defense against bacterial infection, while diquat induces renal injury in Gsr hypomorphic mice. In flies, overexpression of GSR extended lifespan under hyperoxia. In the current study, we investigated the roles of GSR in cochlear antioxidant defense using Gsr homozygous knockout mice that were backcrossed onto the CBA/CaJ mouse strain, a normal-hearing strain that does not carry a specific Cdh23 mutation that causes progressive hair cell degeneration and early onset of hearing loss. Gsr-/- mice displayed a significant decrease in GSR activity and GSH/GSSG ratios in the cytosol of the inner ears. However, Gsr deficiency did not affect ABR (auditory brainstem response) hearing thresholds, wave I amplitudes or wave I latencies in young mice. No histological abnormalities were observed in the cochlea of Gsr-/- mice. Furthermore, there were no differences in the activities of cytosolic glutathione-related enzymes, including glutathione peroxidase and glutamate-cysteine ligase, or the levels of oxidative damage markers in the inner ears between WT and Gsr-/- mice. In contrast, Gsr deficiency resulted in increased activities of cytosolic thioredoxin and thioredoxin reductase in the inner ears. Therefore, under normal physiological conditions, GSR is not essential for the maintenance of antioxidant defenses in mouse cochlea. Given that the thioredoxin system is known to reduce GSSG to GSH in multiple species, our findings suggest that the thioredoxin system can support GSSG reduction in the mouse peripheral auditory system.
谷胱甘肽还原酶(GSR)是谷胱甘肽抗氧化防御系统的关键成员,它将氧化型谷胱甘肽(GSSG)转化为还原型谷胱甘肽(GSH),并维持细胞内谷胱甘肽的氧化还原状态,以保护细胞免受氧化损伤。先前的报道表明,Gsr基因缺失会导致宿主抵抗细菌感染的能力出现缺陷,而百草枯会在Gsr基因低表达小鼠中诱发肾损伤。在果蝇中,GSR的过表达可延长高氧环境下的寿命。在本研究中,我们使用回交至CBA/CaJ小鼠品系的Gsr纯合敲除小鼠,研究了GSR在耳蜗抗氧化防御中的作用,CBA/CaJ小鼠品系是一种听力正常的品系,不携带导致进行性毛细胞退化和早发性听力损失的特定Cdh23突变。Gsr-/-小鼠内耳细胞质中的GSR活性和GSH/GSSG比值显著降低。然而,Gsr基因缺失并未影响幼鼠的听性脑干反应(ABR)听力阈值、I波振幅或I波潜伏期。在Gsr-/-小鼠的耳蜗中未观察到组织学异常。此外,野生型(WT)小鼠和Gsr-/-小鼠内耳中细胞质谷胱甘肽相关酶(包括谷胱甘肽过氧化物酶和谷氨酸-半胱氨酸连接酶)的活性以及氧化损伤标志物的水平没有差异。相反,Gsr基因缺失导致内耳中细胞质硫氧还蛋白和硫氧还蛋白还原酶的活性增加。因此,在正常生理条件下,GSR对于维持小鼠耳蜗的抗氧化防御并非必不可少。鉴于已知硫氧还蛋白系统在多个物种中将GSSG还原为GSH,我们的研究结果表明,硫氧还蛋白系统可以在小鼠外周听觉系统中支持GSSG的还原。