Meniscus cell lysate induces mitochondrial dysfunction of fibroblast-like synoviocytes via upregulating ANT3 in osteoarthritis: aNT3 is a potentially important novel mediator and drug target in osteoarthritis
Meniscus cell lysate induces mitochondrial dysfunction of fibroblast-like synoviocytes via upregulating ANT3 in osteoarthritis: aNT3 is a potentially important novel mediator and drug target in osteoarthritis
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DOI：
10.1302/2046-3758.124.bjr-2022-0135.r2
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发表时间：
2023-04-11
影响因子：
4.6
通讯作者：
Zhang, Z.
中科院分区：
文献类型：
--
作者：
Du, X.;Jiang, Z.;Fang, G.;Liu, R.;Wen, X.;Wu, Y.;Hu, S.;Zhang, Z.
关键词：
本研究旨在探讨半月板细胞裂解液（MCL）在成纤维细胞样滑膜细胞（FLSs）和骨关节炎（OA）中的作用和机制。收集14例患有和不患有OA的患者的半月板和滑膜组织。提取MCL和FLS蛋白，采用液相色谱-质谱（LC-MS）分析。采用酶联免疫吸附试验（ELISA）、流式细胞术、免疫荧光和透射电镜检测MCL和腺嘌呤核苷酸转位酶3 （ANT3）在FLSs中的作用。通过组织学分析确定ANT3在雄性小鼠模型中的表达水平。我们首次发现MCL在OA患者的滑液中大量富集，并通过MCL吞噬促进FLSs中炎症细胞因子的释放。通过LC-MS鉴定并确定ANT3在MCL和OA-FLSs中显著上调，对应于OA-FLSs线粒体功能和细胞活力受损。抑制ANT3可恢复线粒体稳态，从而减轻滑膜炎症。此外，ANT3水平升高抑制ERK磷酸化。具体来说，在mcl诱导的滑膜炎症中，沉默ANT3阻止了ERK磷酸化的抑制，并显著降低了活性氧（ROS）和JC1膜电位的升高。本研究揭示了MCL和ANT3在FLS线粒体中的重要作用。沉默ANT3可以挽救ERK磷酸化，从而恢复FLSs中的线粒体稳态，减轻滑膜炎和OA的发展，为治疗滑膜炎和预防早期OA提供了潜在的靶点。本文引自：骨与关节，2023;12(4):274-284。
This study aimed to investigate the role and mechanism of meniscal cell lysate (MCL) in fibroblast-like synoviocytes (FLSs) and osteoarthritis (OA). Meniscus and synovial tissue were collected from 14 patients with and without OA. MCL and FLS proteins were extracted and analyzed by liquid chromatography‒mass spectrometry (LC‒MS). The roles of MCL and adenine nucleotide translocase 3 (ANT3) in FLSs were examined by enzyme-linked immunosorbent assay (ELISA), flow cytometry, immunofluorescence, and transmission electron microscopy. Histological analysis was performed to determine ANT3 expression levels in a male mouse model. We discovered for the first time that MCL was substantially enriched in the synovial fluid of OA patients and promoted the release of inflammatory cytokines from FLSs through MCL phagocytosis. Through LC‒MS, ANT3 was identified and determined to be significantly upregulated in MCL and OA-FLSs, corresponding to impaired mitochondrial function and cell viability in OA-FLSs. Mitochondrial homeostasis was restored by ANT3 suppression, thereby alleviating synovial inflammation. Furthermore, elevated ANT3 levels inhibited ERK phosphorylation. Specifically, silencing ANT3 prevented inhibition of ERK phosphorylation and significantly reduced the elevation of reactive oxygen species (ROS) and JC1 membrane potential in MCL-induced synovial inflammation. This study revealed the important roles of MCL and ANT3 in FLS mitochondria. Silencing ANT3 rescued ERK phosphorylation, thereby restoring mitochondrial homeostasis in FLSs and alleviating synovitis and OA development, offering a potential target for treating synovitis and preventing early-stage OA. Cite this article: Bone Joint Res 2023;12(4):274–284.