Deubiquitinase USP5 regulates RIPK1 driven pyroptosis in response to myocardial ischemic reperfusion injury.

IF 8.2 2区生物学 Q1 CELL BIOLOGY Cell Communication and Signaling Pub Date : 2024-09-30 DOI:10.1186/s12964-024-01853-x

Wenjing Sun, Hongquan Lu, Lingkun Ma, Cong Ding, Hailan Wang, Yingjie Chu

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Abstract

Background: Gasdermin D (GSDMD) mediated pyroptosis plays a significant role in the pathophysiology of myocardial ischemia/reperfusion (I/R) injury. However, the precise mechanisms regulating pyroptosis remain unclear. In the study, we aimed to investigate the underlying mechanism of pyroptosis in myocardial I/R injury.

Methods: In the present study, we analyzed the effects of USP5 on the RIPK1 kinase activity mediated pyroptosis in vitro after H/R (hypoxia/reoxygenation) and in vivo in a MI/R mouse model. TTC and Evan's blue dye, Thioflavin S and immunohistochemistry staining were performed in wild-type, RIPK1^flox/flox Cdh5-Cre and USP5 deficiency mice. CMEC cells were transfected with si-USP5. HEK293T cells were transfected with USP5 and RIPK1 overexpression plasmid or its mutants. The levels of USP5, RIPK1, Caspase-8, FADD and GSDMD were determined by Western blot. Protein interactions were evaluated by immunoprecipitation. The protein colocalization in cells was monitored using a confocal microscope.

Results: In this study, our data demonstrate that RIPK1 is essential for limiting cardiac endothelial cell (CMEC) pyroptosis mediated by caspase-8 in response to myocardial I/R. Additionally, we investigate the role of ubiquitin-specific protease 5 (USP5) as a deubiquitinase for RIPK1. Mechanistically, USP5 interacts with RIPK1, leading to its deubiquitination and stabilization.

Conclusions: These findings offer new insights into the role of USP5 in regulating RIPK1-induced pyroptosis.

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去泛素化酶 USP5 在心肌缺血再灌注损伤时调控 RIPK1 驱动的热蛋白沉积。

背景：Gasdermin D（GSDMD）介导的热蛋白沉积在心肌缺血/再灌注（I/R）损伤的病理生理学中起着重要作用。然而，调控热蛋白沉积的确切机制仍不清楚。本研究旨在探讨心肌I/R损伤中热蛋白沉积的内在机制：在本研究中，我们分析了 USP5 在 H/R（缺氧/复氧）后体外和 MI/R 小鼠模型体内对 RIPK1 激酶活性介导的热凋亡的影响。在野生型小鼠、RIPK1flox/flox Cdh5-Cre小鼠和USP5缺乏小鼠中进行了TTC和伊文氏蓝染料、硫黄素S和免疫组织化学染色。用 si-USP5 转染 CMEC 细胞。用 USP5 和 RIPK1 过表达质粒或其突变体转染 HEK293T 细胞。用 Western 印迹法测定 USP5、RIPK1、Caspase-8、FADD 和 GSDMD 的水平。蛋白质相互作用通过免疫沉淀法进行评估。使用共聚焦显微镜监测细胞中的蛋白质共聚焦：在这项研究中，我们的数据证明了 RIPK1 对于限制心肌内皮细胞（CMEC）在应对心肌 I/R 时由 caspase-8 介导的热凋亡至关重要。此外，我们还研究了泛素特异性蛋白酶 5（USP5）作为 RIPK1 的去泛素化酶的作用。从机制上讲，USP5 与 RIPK1 相互作用，导致其去泛素化和稳定化：这些发现为了解 USP5 在调节 RIPK1 诱导的化脓过程中的作用提供了新的视角。

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

Cell Communication and Signaling CELL BIOLOGY-

CiteScore

11.00

自引率

0.00%

发文量

180

期刊介绍： Cell Communication and Signaling (CCS) is a peer-reviewed, open-access scientific journal that focuses on cellular signaling pathways in both normal and pathological conditions. It publishes original research, reviews, and commentaries, welcoming studies that utilize molecular, morphological, biochemical, structural, and cell biology approaches. CCS also encourages interdisciplinary work and innovative models, including in silico, in vitro, and in vivo approaches, to facilitate investigations of cell signaling pathways, networks, and behavior. Starting from January 2019, CCS is proud to announce its affiliation with the International Cell Death Society. The journal now encourages submissions covering all aspects of cell death, including apoptotic and non-apoptotic mechanisms, cell death in model systems, autophagy, clearance of dying cells, and the immunological and pathological consequences of dying cells in the tissue microenvironment.