Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway

As a pattern recognition receptor, Toll-like receptor 4 (TLR4) is crucial for the development and progression of acute kidney injury (AKI). This study aims to explore whether the deubiquitinase Usp9x influences the TLR4/NF-B pathway to cause sepsis-induced acute kidney injury (S-AKI). The model of A...

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Main Authors: Shuhao Gong, Huawei Xiong, Yingchao Lei, Shipeng Huang, Yingdong Ouyang, Chunshui Cao, Ying Wang
Format: Article
Language:English
Published: Taylor & Francis Group 2024-12-01
Series:Renal Failure
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Online Access:https://www.tandfonline.com/doi/10.1080/0886022X.2024.2361089
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author Shuhao Gong
Huawei Xiong
Yingchao Lei
Shipeng Huang
Yingdong Ouyang
Chunshui Cao
Ying Wang
author_facet Shuhao Gong
Huawei Xiong
Yingchao Lei
Shipeng Huang
Yingdong Ouyang
Chunshui Cao
Ying Wang
author_sort Shuhao Gong
collection DOAJ
description As a pattern recognition receptor, Toll-like receptor 4 (TLR4) is crucial for the development and progression of acute kidney injury (AKI). This study aims to explore whether the deubiquitinase Usp9x influences the TLR4/NF-B pathway to cause sepsis-induced acute kidney injury (S-AKI). The model of AKI was established in Sprague-Dawley rats using the cecal ligation and puncture (CLP) method, while renal tubular epithelial cell NRK-52E was stimulated with lipopolysaccharide (LPS) in vitro. All plasmids were transfected into NRK-52E cells according to the indicated group. The deubiquitinase of TLR4 was predicted by the online prediction software Ubibrowser. Subsequently, Western blot and Pearson correlation analysis identified Usp9x protein as a potential candidate. Co-IP analysis verified the interaction between TLR4 and Usp9x. Further research revealed that overexpression of Usp9x inhibited degradation of TLR4 protein by downregulating its ubiquitination modification levels. Both in vivo and in vitro experiments observed that interference with Usp9x effectively alleviated the inflammatory response and apoptosis of renal tubular epithelial cells (RTECs) induced by CLP or LPS, whereas overexpression of TLR4 reversed this situation. Transfection with sh-Usp9x in NRK-52E cells suppressed the expression of proteins associated with the TLR4/NF-κB pathway induced by LPS. Moreover, the overexpression of TLR4 reversed the effect of sh-Usp9x transfection. Therefore, the deubiquitinase Usp9x interacts with TLR4, leading to the upregulation of its expression through deubiquitination modification, and the activation of the TLR4/NF-κB signaling pathway, thereby promoting inflammation and apoptosis in renal tubular epithelial cells and contributing to sepsis-induced acute kidney injury.
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issn 0886-022X
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publishDate 2024-12-01
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series Renal Failure
spelling doaj-art-068fbd70e5c8476bb1473e968788499a2025-08-20T02:29:58ZengTaylor & Francis GroupRenal Failure0886-022X1525-60492024-12-0146210.1080/0886022X.2024.2361089Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathwayShuhao Gong0Huawei Xiong1Yingchao Lei2Shipeng Huang3Yingdong Ouyang4Chunshui Cao5Ying Wang6Department of Emergency, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaDepartment of Emergency, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaDepartment of Emergency, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaDepartment of Emergency, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaDepartment of Emergency, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaDepartment of Emergency, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaDepartment of Nephrology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, ChinaAs a pattern recognition receptor, Toll-like receptor 4 (TLR4) is crucial for the development and progression of acute kidney injury (AKI). This study aims to explore whether the deubiquitinase Usp9x influences the TLR4/NF-B pathway to cause sepsis-induced acute kidney injury (S-AKI). The model of AKI was established in Sprague-Dawley rats using the cecal ligation and puncture (CLP) method, while renal tubular epithelial cell NRK-52E was stimulated with lipopolysaccharide (LPS) in vitro. All plasmids were transfected into NRK-52E cells according to the indicated group. The deubiquitinase of TLR4 was predicted by the online prediction software Ubibrowser. Subsequently, Western blot and Pearson correlation analysis identified Usp9x protein as a potential candidate. Co-IP analysis verified the interaction between TLR4 and Usp9x. Further research revealed that overexpression of Usp9x inhibited degradation of TLR4 protein by downregulating its ubiquitination modification levels. Both in vivo and in vitro experiments observed that interference with Usp9x effectively alleviated the inflammatory response and apoptosis of renal tubular epithelial cells (RTECs) induced by CLP or LPS, whereas overexpression of TLR4 reversed this situation. Transfection with sh-Usp9x in NRK-52E cells suppressed the expression of proteins associated with the TLR4/NF-κB pathway induced by LPS. Moreover, the overexpression of TLR4 reversed the effect of sh-Usp9x transfection. Therefore, the deubiquitinase Usp9x interacts with TLR4, leading to the upregulation of its expression through deubiquitination modification, and the activation of the TLR4/NF-κB signaling pathway, thereby promoting inflammation and apoptosis in renal tubular epithelial cells and contributing to sepsis-induced acute kidney injury.https://www.tandfonline.com/doi/10.1080/0886022X.2024.2361089Sepsis-induced acute kidney injuryubiquitination modificationinflammationapoptosisUsp9xTLR4/NF-κB
spellingShingle Shuhao Gong
Huawei Xiong
Yingchao Lei
Shipeng Huang
Yingdong Ouyang
Chunshui Cao
Ying Wang
Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway
Renal Failure
Sepsis-induced acute kidney injury
ubiquitination modification
inflammation
apoptosis
Usp9x
TLR4/NF-κB
title Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway
title_full Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway
title_fullStr Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway
title_full_unstemmed Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway
title_short Usp9x contributes to the development of sepsis-induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the TLR4/nf-κb pathway
title_sort usp9x contributes to the development of sepsis induced acute kidney injury by promoting inflammation and apoptosis in renal tubular epithelial cells via activation of the tlr4 nf κb pathway
topic Sepsis-induced acute kidney injury
ubiquitination modification
inflammation
apoptosis
Usp9x
TLR4/NF-κB
url https://www.tandfonline.com/doi/10.1080/0886022X.2024.2361089
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