HSF1 Increases EOGT-Mediated Glycosylation of Notch1 to Promote IL-1β-Induced Inflammatory Injury of Chondrocytes.

IF 2.7 4区医学 Q1 ORTHOPEDICS

CARTILAGE Pub Date : 2024-02-16 DOI:10.1177/19476035241229211

Yuanchi Huang, Wenjie Pan, Huanli Bao, Xiangxiang Sun, Chao Xu, Jianbing Ma

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引用次数: 0

Abstract

Objective: Osteoarthritis (OA) is the most common arthritic disease in humans. Nevertheless, the pathogenic mechanism of OA remains unclear. This study aimed to explore that heat-shock transcription factor 1 (HSF1) facilitated interleukin-1 beta (IL-1β) chondrocyte injury by increasing Notch1 O-linked N-acetylglucosamine (O-GlcNAc) modification level.

Design: Human chondrocytes were incubated with 5 ng/ml interleukin-1 beta (IL-1β) for 24 h to establish OA cell model. The messenger RNA (mRNA) or protein expressions were assessed using reverse transcription-quantitative polymerase chain reaction, western blot, or immunofluorescence. Chondrocyte viability was examined by Cell Counting Kit-8 assay. Enzyme-linked immunosorbent assay was employed to detect the secretion levels of interleukin-6 (IL-6) and interleukin-8 (IL-8). Immunoprecipitation was adopted to detect Notch1 O-GlcNAc modification level. The interaction between HSF1 and epidermal growth factor-like (EGF) domain-specific O-GlcNAc transferase (EOGT) promoter was analyzed by dual-luciferase reporter gene and chromatin immunoprecipitation assays.

Results: Herein, our results demonstrated that HSF1, EOGT, Notch1, and Notch1 intracellular domain (NICD1) expressions in chondrocytes were markedly increased by IL-1β stimulation. EOGT elevated Notch1 expression in IL-1β-treated chondrocytes by increasing Notch1 O-GlcNAc modification level. EOGT silencing reduced IL-1β-induced chondrocyte inflammatory injury. In addition, HSF1 knockdown relieved IL-1β-induced chondrocyte inflammatory injury. Molecular interaction experiment proved that HSF1 transcriptionally activated EOGT expression in IL-1β-treated chondrocytes.

Conclusions: HSF1 promoted IL-1β-induced inflammatory injury in chondrocytes by increasing EOGT-mediated glycosylation of Notch1.

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HSF1 可增加 EOGT 介导的 Notch1 糖基化，从而促进 IL-1β 诱导的软骨细胞炎症损伤。

目的：骨关节炎（OA）是人类最常见的关节炎疾病：骨关节炎（OA）是人类最常见的关节炎疾病。然而，OA 的致病机制仍不清楚。本研究旨在探讨热休克转录因子1（HSF1）通过增加Notch1 O-连接的N-乙酰葡糖胺（O-GlcNAc）修饰水平促进白细胞介素-1β（IL-1β）软骨细胞损伤：设计：将人软骨细胞与 5 ng/ml 的白细胞介素-1β（IL-1β）培养 24 小时，以建立 OA 细胞模型。用逆转录-定量聚合酶链反应、Western 印迹或免疫荧光法评估信使 RNA（mRNA）或蛋白质的表达。用细胞计数试剂盒-8检测软骨细胞的活力。酶联免疫吸附法检测白细胞介素-6（IL-6）和白细胞介素-8（IL-8）的分泌水平。免疫沉淀法检测 Notch1 O-GlcNAc 修饰水平。通过双荧光素酶报告基因和染色质免疫沉淀实验分析了HSF1与表皮生长因子样（EGF）结构域特异性O-GlcNAc转移酶（EOGT）启动子之间的相互作用：结果：我们的研究结果表明，HSF1、EOGT、Notch1和Notch1胞内结构域（NICD1）在软骨细胞中的表达在IL-1β刺激下显著增加。EOGT通过增加Notch1 O-GlcNAc修饰水平提高了Notch1在IL-1β处理的软骨细胞中的表达。沉默EOGT可减少IL-1β诱导的软骨细胞炎症损伤。此外，HSF1敲除可缓解IL-1β诱导的软骨细胞炎症损伤。分子相互作用实验证明，HSF1转录激活了IL-1β处理的软骨细胞中EOGT的表达：结论：HSF1通过增加EOGT介导的Notch1糖基化促进了IL-1β诱导的软骨细胞炎症损伤。

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

CARTILAGE ORTHOPEDICS-

CiteScore

6.90

自引率

7.10%

发文量

期刊介绍： CARTILAGE publishes articles related to the musculoskeletal system with particular attention to cartilage repair, development, function, degeneration, transplantation, and rehabilitation. The journal is a forum for the exchange of ideas for the many types of researchers and clinicians involved in cartilage biology and repair. A primary objective of CARTILAGE is to foster the cross-fertilization of the findings between clinical and basic sciences throughout the various disciplines involved in cartilage repair. The journal publishes full length original manuscripts on all types of cartilage including articular, nasal, auricular, tracheal/bronchial, and intervertebral disc fibrocartilage. Manuscripts on clinical and laboratory research are welcome. Review articles, editorials, and letters are also encouraged. The ICRS envisages CARTILAGE as a forum for the exchange of knowledge among clinicians, scientists, patients, and researchers. The International Cartilage Repair Society (ICRS) is dedicated to promotion, encouragement, and distribution of fundamental and applied research of cartilage in order to permit a better knowledge of function and dysfunction of articular cartilage and its repair.