Overexpression of DTX1 inhibits D-GalN/TNF-α-induced pyroptosis and inflammation in hepatocytes by regulating NLRP3 ubiquitination.

IF 2.2 4区医学 Q3 TOXICOLOGY

Toxicology Research Pub Date : 2024-09-23 eCollection Date: 2024-10-01 DOI:10.1093/toxres/tfae145

Mingshui Liu, Jing Gu, Li Chen, Wei Sun, Xiaoping Huang, Jianhe Gan

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引用次数: 0

Abstract

Background: Acute liver injury (ALI) is characterized by massive hepatocyte death and has high mortality and poor prognosis. Hepatocyte pyroptosis plays a key role in the pathophysiology of ALI and is involved in the inflammatory response mediated by NOD-like receptor protein 3 (NLRP3) inflammasome activation. Deltex 1 (DTX1) is a single transmembrane protein with ubiquitin E3 ligase activity and is closely involved in cell growth, differentiation, and apoptosis, as well as intracellular signal transduction. However, little is known about the influence of DTX1 on ALI. This study aimed to investigate the role of DTX1 in pyroptosis and inflammation induced by D-galactosamine (D-GalN) and tumor necrosis factoralpha (TNF-α) in human hepatocytes (LO2 cells) in vitro.

Methods: Cell pyroptosis was measured by flow cytometry. The levels of DTX1, pyroptosis-associated proteins, and inflammatory cytokines were detected by quantitative real-time polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay. Immunofluorescence staining, co-immunoprecipitation, ubiquitination, and luciferase reporter and chromatin immunoprecipitation assays were performed to detect the regulation between DTX1 and NLRP3 or hepatocyte nuclear factor 4 alpha (HNF4α). Analysis of variance was performed to compare groups.

Results: We found that DTX1 was decreased in D-GalN/TNF-α-induced LO2 cells. DTX1 overexpression significantly inhibited D-GalN/TNF-α-induced cell pyroptosis and inflammation. DTX1 interacted with NLRP3 and induced NLRP3 ubiquitination and degradation. Furthermore, by targeting NLRP3, DTX1 knockdown significantly induced cell pyroptosis and inflammation. In addition, HNF4α promoted DTX1 transcription by binding with its promoter.

Conclusion: Our study revealed that DTX1 suppressed D-GalN/TNF-α-induced hepatocyte pyroptosis and inflammation by regulating NLRP3 ubiquitination.

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通过调节 NLRP3 泛素化，过表达 DTX1 可抑制 D-GalN/TNF-α 诱导的肝细胞脓毒症和炎症。

背景：急性肝损伤（ALI）的特点是肝细胞大量死亡，死亡率高，预后差。肝细胞热解在 ALI 的病理生理学中起着关键作用，并参与了由 NOD 样受体蛋白 3（NLRP3）炎性基因组激活介导的炎症反应。Deltex 1（DTX1）是一种具有泛素 E3 连接酶活性的单跨膜蛋白，密切参与细胞生长、分化和凋亡以及细胞内信号转导。然而，人们对 DTX1 对 ALI 的影响知之甚少。本研究旨在探讨 DTX1 在体外由 D-半乳糖胺（D-GalN）和肿瘤坏死因子α（TNF-α）诱导的人肝细胞（LO2 细胞）热休克和炎症中的作用：方法：采用流式细胞术测量细胞的热解过程。方法：采用流式细胞仪测量细胞热解，通过实时定量聚合酶链式反应、Western 印迹和酶联免疫吸附试验检测 DTX1、热解相关蛋白和炎性细胞因子的水平。通过免疫荧光染色、共免疫沉淀、泛素化、荧光素酶报告和染色质免疫沉淀检测 DTX1 与 NLRP3 或肝细胞核因子 4 alpha（HNF4α）之间的调控。对各组进行方差分析比较：结果：我们发现，D-GalN/TNF-α诱导的LO2细胞中DTX1减少。DTX1的过表达能显著抑制D-GalN/TNF-α诱导的细胞脓毒症和炎症。DTX1 与 NLRP3 相互作用，诱导 NLRP3 泛素化和降解。此外，通过靶向 NLRP3，敲除 DTX1 能显著诱导细胞裂解和炎症。此外，HNF4α通过与其启动子结合促进了DTX1的转录：我们的研究发现，DTX1 通过调节 NLRP3 泛素化抑制了 D-GalN/TNF-α 诱导的肝细胞化脓和炎症。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Toxicology Research TOXICOLOGY-

CiteScore

3.60

自引率

0.00%

发文量

期刊介绍： A multi-disciplinary journal covering the best research in both fundamental and applied aspects of toxicology