去泛素化酶 USP19 通过靶向 NOX4 蛋白调节 SH-SY5Y 细胞中的铁死亡和线粒体损伤。

Deubiquitinating Enzyme USP19 Regulates Ferroptosis and Mitochondrial Damage in SH-SY5Y Cells by Targeting the NOX4 Protein.

机构信息

College of Integrated Traditional Chinese and Western Medicine, Fujian University of Traditional Chinese Medicine, Fu Zhou, China.

Department of Neurosurgery, Fuzhou Second Hospital, Fu Zhou, China.

出版信息

J Alzheimers Dis. 2024;100(3):799-808. doi: 10.3233/JAD-231193.

DOI:10.3233/JAD-231193

PMID:38943386

Abstract

BACKGROUND

Ferroptosis is extremely relevant to the progression of neurodegenerative pathologies such as Alzheimer's disease (AD). Ubiquitin-specific proteases (USP) can affect the NADPH oxidase family.

OBJECTIVE

Our study aimed to elucidate the potential role and molecular basis of a certain USP19 in reducing ferroptosis and mitochondrial injury in AD cells by targeting NOX4 stability.

METHODS

The deubiquitinase USP family gene USP19, which affects the stability of NOX4 protein, was first screened. The cell model of AD was constructed after interfering with SH-SY5Y cells by Aβ1-40, and then SH-SY5Y cells were infected with lentiviral vectors to knock down USP19 and overexpress NOX4, respectively. Finally, the groups were tested for cell viability, changes in cellular mitochondrial membrane potential, lipid reactive oxygen species, intracellular iron metabolism, and NOX4, Mf1, Mf2, and Drp1 protein expression.

RESULTS

5 μmol/L Aβ1-40 intervened in SH-SY5Y cells for 24 h to construct a cell model of AD. Knockdown of USP19 decreased the expression of NOX4 protein, promoted the expression of mitochondrial fusion proteins Mnf1 and Mnf2, and inhibited the expression of the splitting protein Drp1. Furthermore, USP19 knockdown decreased mitochondrial membrane potential, SOD, MDA, intracellular iron content and increased GSH/GSSG ratio in SH-SY5Y cells. Our study revealed that NOX4 protein interacts with USP19 and knockdown of USP19 enhanced ubiquitination to maintain NOX4 protein stability.

CONCLUSIONS

USP19 attenuates mitochondrial damage in SH-SY5Y cells by targeting NOX4 protein with Aβ1-40.

摘要

背景

铁死亡与阿尔茨海默病（AD）等神经退行性病变的进展密切相关。泛素特异性蛋白酶（USP）可以影响 NADPH 氧化酶家族。

目的

本研究旨在通过靶向 NOX4 稳定性，阐明特定 USP19 在降低 AD 细胞中铁死亡和线粒体损伤方面的潜在作用和分子基础。

方法

首先筛选影响 NOX4 蛋白稳定性的去泛素化酶 USP 家族基因 USP19。用 Aβ1-40 干扰 SH-SY5Y 细胞构建 AD 细胞模型，然后用慢病毒载体感染 SH-SY5Y 细胞分别敲低 USP19 和过表达 NOX4，最后检测细胞活力、细胞线粒体膜电位变化、脂质活性氧、细胞内铁代谢以及 NOX4、Mf1、Mf2 和 Drp1 蛋白表达。

结果

5 μmol/L Aβ1-40 干预 SH-SY5Y 细胞 24 h 构建 AD 细胞模型。USP19 敲低降低了 NOX4 蛋白的表达，促进了线粒体融合蛋白 Mnf1 和 Mnf2 的表达，抑制了分裂蛋白 Drp1 的表达。此外，USP19 敲低降低了 SH-SY5Y 细胞的线粒体膜电位、SOD、MDA、细胞内铁含量，增加了 GSH/GSSG 比值。本研究表明，NOX4 蛋白与 USP19 相互作用，USP19 敲低增强了泛素化以维持 NOX4 蛋白稳定性。