去泛素化酶 USP16 通过 Drp1 依赖性线粒体分裂和 NLRP3 炎性体激活诱导痛风性关节炎。
Deubiquitinase USP16 induces gouty arthritis via Drp1-dependent mitochondrial fission and NLRP3 inflammasome activation.
机构信息
Key Laboratory of Microecology-Immune Regulatory Network and Related Diseases, School of Basic Medicine, Jiamusi University, Heilongjiang Province, Jiamusi, 154000, People's Republic of China.
出版信息
Arthritis Res Ther. 2023 Jul 24;25(1):126. doi: 10.1186/s13075-023-03095-7.
BACKGROUND
Gouty arthritis is the most frequently diagnosed inflammatory arthritis worldwide. Dynamin-related protein 1 (Drp1), a regulator of mitochondrial fission, contributes to various inflammatory disorders via activating NLRP3 inflammasome. However, the biological role of Drp1 in gouty arthritis remains undefined.
METHODS
A mouse model of monosodium urate (MSU)-induced gouty arthritis and MSU-stimulated macrophages were established as in vivo and in vitro models, respectively. Histological changes were assessed by H&E and IHC analysis. RT-qPCR and western blot were used to detect the expression of Drp1 and the key molecules in joint tissues and macrophages. Cytokine secretion was measured by ELISA assay, and antioxidant enzymes activities and LDH release were monitored using commercial kits. Mitochondrial structure and functions were assessed by transmission electron microscopy (TEM) and MitoSOX staining. Co-IP and GST pull-down assay were used to detect the direct interaction between USP16 and Drp1, as well as the ubiquitination of Drp1.
RESULTS
Drp1 was elevated in MSU-induced gouty arthritis model, and it induced gouty arthritis via NF-κB pathway and NLRP3 inflammasome activation. In addition, Drp1 activated NF-κB/NLRP3 signaling via modulating mitochondrial fission. Mechanistically, USP16 mediated deubiquitination and stabilization of Drp1 through its direct interaction with Drp1. Functional studies further showed that USP16 was highly expressed in MSU-stimulated macrophages and induced gouty arthritis via Drp1-dependent NLRP3 inflammasome activation.
CONCLUSION
Deubiquitinase USP16 induced gouty arthritis via Drp1-dependent mitochondrial fission and NF-κB/NLRP3 signaling.
背景
痛风性关节炎是全球最常见的炎性关节炎。动力相关蛋白 1（Drp1）是一种线粒体分裂的调节蛋白，通过激活 NLRP3 炎性小体参与各种炎症性疾病。然而，Drp1 在痛风性关节炎中的生物学作用尚不清楚。
方法
建立了尿酸单钠（MSU）诱导的痛风性关节炎小鼠模型和 MSU 刺激的巨噬细胞体外模型，分别作为体内和体外模型。通过 H&E 和 IHC 分析评估组织学变化。使用 RT-qPCR 和 Western blot 检测关节组织和巨噬细胞中 Drp1 及关键分子的表达。通过 ELISA 测定细胞因子分泌，通过商业试剂盒监测抗氧化酶活性和 LDH 释放。使用透射电子显微镜（TEM）和 MitoSOX 染色评估线粒体结构和功能。通过 Co-IP 和 GST 下拉实验检测 USP16 和 Drp1 之间的直接相互作用，以及 Drp1 的泛素化。
结果
Drp1 在 MSU 诱导的痛风性关节炎模型中上调，并通过 NF-κB 途径和 NLRP3 炎性小体激活诱导痛风性关节炎。此外，Drp1 通过调节线粒体分裂激活 NF-κB/NLRP3 信号。机制上，USP16 通过与 Drp1 的直接相互作用介导 Drp1 的去泛素化和稳定。功能研究进一步表明，USP16 在 MSU 刺激的巨噬细胞中高表达，并通过 Drp1 依赖性 NLRP3 炎性小体激活诱导痛风性关节炎。
结论
去泛素酶 USP16 通过 Drp1 依赖性线粒体分裂和 NF-κB/NLRP3 信号诱导痛风性关节炎。
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