GGT1 Suppresses the Development of Ferroptosis and Autophagy in Mouse Retinal Ganglion Cell Through Targeting GCLC.

IF 3.1 Q1 OPHTHALMOLOGY
Eye and Brain Pub Date : 2023-11-21 eCollection Date: 2023-01-01 DOI:10.2147/EB.S434280
Guihua Xu, Juanjuan Wang, Yiting Zhang, Zilin Chen, Ruidong Deng
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引用次数: 0

Abstract

Background: Glaucoma is a neurodegenerative disorder characterized with optic nerve injury and the loss of retinal ganglion cells (RGCs). Ferroptosis has been proved to be associated with the degradation of RGCs. The aim of this study is to elucidate the relationship between ferroptosis and glaucoma pathogenesis, and unveil the underlying mechanism.

Methods: Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the proliferation of RGCs. The accumulation of cellular iron was measured by Iron assay kit, and the level of reactive oxygen species (ROS) was detected by fluorescence probe. The mitochondrial morphology and autophagosomes were analysed by using transmission electron microscopy (TEM). The contents of glutathione (GSH) and malondialdehyde (MDA) were tested by a GSH assay kit and an MDA detection kit, respectively. The expression of autophagy-related proteins was detected by Western blotting.

Results: A serious cell damage, aberrant iron homeostasis, and oxidative stress was shown in RGC-5 after oxygen-glucose deprivation/reoxygenation (OGD/R) treatment and gamma-Glutamyl transpeptidase 1 (GGT1) knockdown, but these effects were significantly alleviated by overexpression of GGT1 or ferroptosis inhibitors. The TEM and immunofluorescent results indicated that mitochondria impairment and autophagosome accumulation in OGD/R-treated cells was improved after GGT1 overexpression, while the phenomenon in GGT1-silenced cells was aggravated. Furthermore, we found that GGT1 can interact with glutamate cysteine ligase catalytic subunit (GCLC) to inhibit autophagy and ferroptosis in RGC-5 cells.

Conclusion: GGT1 represses autophagy in RGC-5 cells by targeting GCLC, which further restrains the development of ferroptosis in cells.

GGT1通过靶向GCLC抑制小鼠视网膜神经节细胞铁下垂和自噬的发生。
背景:青光眼是一种以视神经损伤和视网膜神经节细胞(RGCs)丧失为特征的神经退行性疾病。铁下垂已被证明与rgc的降解有关。本研究的目的是阐明铁下垂与青光眼发病的关系,并揭示其潜在的机制。方法:采用甲基噻唑四氮唑(MTT)法检测RGCs的增殖情况。采用铁含量测定试剂盒检测细胞铁积累,荧光探针检测活性氧(ROS)水平。透射电镜观察了线粒体形态和自噬体的变化。分别用谷胱甘肽(GSH)测定试剂盒和丙二醛(MDA)测定试剂盒检测各组血清谷胱甘肽(GSH)和丙二醛(MDA)含量。Western blotting检测自噬相关蛋白的表达。结果:在氧-葡萄糖剥夺/再氧化(OGD/R)治疗和γ -谷氨酰转肽酶1 (GGT1)敲低后,RGC-5出现严重的细胞损伤、铁稳态异常和氧化应激,但过表达GGT1或铁凋亡抑制剂可显著减轻这些影响。透射电镜和免疫荧光结果显示,GGT1过表达后,OGD/ r处理细胞的线粒体损伤和自噬体积累得到改善,而GGT1沉默细胞的这一现象加剧。此外,我们发现GGT1可以与谷氨酸半胱氨酸连接酶催化亚基(GCLC)相互作用,抑制RGC-5细胞的自噬和铁凋亡。结论:GGT1通过靶向GCLC抑制RGC-5细胞的自噬,进一步抑制细胞铁下垂的发生。
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来源期刊
Eye and Brain
Eye and Brain OPHTHALMOLOGY-
CiteScore
7.90
自引率
2.30%
发文量
12
审稿时长
16 weeks
期刊介绍: Eye and Brain is an international, peer-reviewed, open access journal focusing on basic research, clinical findings, and expert reviews in the field of visual science and neuro-ophthalmology. The journal’s unique focus is the link between two well-known visual centres, the eye and the brain, with an emphasis on the importance of such connections. All aspects of clinical and especially basic research on the visual system are addressed within the journal as well as significant future directions in vision research and therapeutic measures. This unique journal focuses on neurological aspects of vision – both physiological and pathological. The scope of the journal spans from the cornea to the associational visual cortex and all the visual centers in between. Topics range from basic biological mechanisms to therapeutic treatment, from simple organisms to humans, and utilizing techniques from molecular biology to behavior. The journal especially welcomes primary research articles or review papers that make the connection between the eye and the brain. Specific areas covered in the journal include: Physiology and pathophysiology of visual centers, Eye movement disorders and strabismus, Cellular, biochemical, and molecular features of the visual system, Structural and functional organization of the eye and of the visual cortex, Metabolic demands of the visual system, Diseases and disorders with neuro-ophthalmic manifestations, Clinical and experimental neuro-ophthalmology and visual system pathologies, Epidemiological studies.
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