胆固醇24-羟化酶CYP46A1的过表达通过调节Nrf2通路减轻视神经挤压损伤中视网膜功能障碍和神经节细胞损失

IF 2.7 2区 医学 Q1 OPHTHALMOLOGY
Zhenli Long , Jun Zhang , Jiazhen Feng, Tao He
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引用次数: 0

摘要

胆固醇24-羟化酶CYP46A1 (CYP46A1)已被证实与多种神经系统疾病的过程相关,但其在神经退行性视神经疾病中的作用尚不清楚。本文旨在探讨CYP46A1对小鼠视网膜神经节细胞(RGCs)和视网膜功能的神经保护作用。玻璃体内注射raav后小鼠视神经损伤(ONC)。视网膜平载免疫荧光染色定量RGCs存活。采用视网膜电图(ERG)和视动反应(OMR)定量评价视网膜电生理功能和视力。采用tdt介导的dUTP镍端标记(TUNEL)染色法定量RGCs的凋亡情况。Western blot检测CYP46A1、b细胞淋巴瘤2 (Bcl-2)、Bcl-2相关X蛋白(Bax)、kelch样ech相关蛋白1 (Keap1)、核因子红系2相关因子(Nrf2)、血红素加氧酶1 (HO1)蛋白表达水平。ONC损伤后,CYP46A1表达增加,RGCs存活率、光负反应幅度(PhNR)、视力下降;a波和b波无明显变化。Western blotting和视网膜冷冻切片染色证实,玻璃体内注射rAAV-CYP46A1-EGFP可上调CYP46A1的表达。CYP46A1过表达可减轻onc诱导的RGCs丢失和视网膜电生理功能障碍。过表达CYP46A1可显著抑制RGCs凋亡,降低Bax表达,增加Bcl-2表达。此外,CYP46A1的上调导致Keap1表达降低,Nrf2和HO-1水平升高。综上所述,我们的研究结果表明,CYP46A1的过表达可以通过激活Nrf2信号通路来防止rgc的丢失,保护rgc的电生理功能。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Overexpression of cholesterol 24-hydroxylase CYP46A1 attenuates retinal dysfunction and ganglion cell loss via regulating the Nrf2 pathway in optic nerve crush injury
Cholesterol 24-hydroxylase CYP46A1 (CYP46A1) has been confirmed to be correlated with the processes of multiple neurological disorders, but its role in neurodegenerative optic diseases remains unclear. This article aimed to evaluate the neuroprotective effects of CYP46A1 on mouse retinal ganglion cells (RGCs) and retinal function. Mice were subjected to optic nerve crush (ONC) injury after intravitreal injection of rAAVs. RGCs' survival was quantified by immunofluorescence staining of retinal flat mounts. Retinal electrophysiological function and visual acuity were quantitatively assessed using electroretinography (ERG) and optomotor response (OMR). The TdT-mediated dUTP nick-end labeling (TUNEL) staining was employed to quantify the apoptosis of RGCs. The protein expression level of CYP46A1, B-cell lymphoma 2 (Bcl-2), BCL-2-associated X protein (Bax), Kelch-like ECH-associated protein 1 (Keap1), nuclear factor erythroid 2-related factor (Nrf2), and heme oxygenase 1 (HO1) were validated by Western blot. After ONC injury, CYP46A1 expression increased, while RGCs’ survival rate, the amplitude of photopic negative response (PhNR), and visual acuity decreased; there were no significant changes in the a-wave and b-wave. Western blotting and retinal cryosection staining confirmed that intravitreal injection of rAAV-CYP46A1-EGFP upregulated CYP46A1 expression. CYP46A1 overexpression mitigated ONC-induced RGCs loss and retinal electrophysiological dysfunction. The overexpression of CYP46A1 could significantly inhibit RGCs apoptosis, reduce Bax expression and increase Bcl-2 expression. Additionally, the upregulation of CYP46A1 led to decreased Keap1 expression and increased Nrf2 and HO-1 levels. In conclusion, our results demonstrated that the overexpression of CYP46A1 could prevent the loss of RGCs and protect the electrophysiological function by activating Nrf2 signaling pathway.
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来源期刊
Experimental eye research
Experimental eye research 医学-眼科学
CiteScore
6.80
自引率
5.90%
发文量
323
审稿时长
66 days
期刊介绍: The primary goal of Experimental Eye Research is to publish original research papers on all aspects of experimental biology of the eye and ocular tissues that seek to define the mechanisms of normal function and/or disease. Studies of ocular tissues that encompass the disciplines of cell biology, developmental biology, genetics, molecular biology, physiology, biochemistry, biophysics, immunology or microbiology are most welcomed. Manuscripts that are purely clinical or in a surgical area of ophthalmology are not appropriate for submission to Experimental Eye Research and if received will be returned without review.
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