Department of Teaching Administration, Xiangyang No. 1 People's Hospital, Hubei University of Medicine, Xiangyang, Hubei 441000, P.R. China.
Department of Radiology, Xiangyang No. 1 People's Hospital, Hubei University of Medicine, Xiangyang, Hubei 441000, P.R. China.
Mol Med Rep. 2024 Aug;30(2). doi: 10.3892/mmr.2024.13273. Epub 2024 Jun 28.
Osteoarthritis (OA) is a chronic disease that involves chondrocyte injury. ADAMTS5 has been confirmed to mediate chondrocyte injury and thus regulate OA progression, but its underlying molecular mechanisms remain unclear. In the present study, interleukin‑1β (IL‑1β)‑induced chondrocytes were used to mimic OA . Cell proliferation and apoptosis were assessed by MTT assay, EdU assay and flow cytometry, and protein levels of ADAMTS5, specificity protein 1 (SP1), matrix‑related markers and Wnt/β‑catenin pathway‑related markers were examined using western blotting. In addition, ELISA was performed to measure the concentrations of inflammation factors, and oxidative stress was evaluated by detecting SOD activity and MDA levels. The mRNA expression levels of ADAMTS5 and SP1 were determined by reverse transcription‑quantitative PCR, and the interaction between SP1 and ADAMTS5 was analyzed using a dual‑luciferase reporter assay and chromatin immunoprecipitation assay. IL‑1β suppressed proliferation, but promoted apoptosis, extracellular matrix degradation, inflammation and oxidative stress in chondrocytes. ADAMTS5 was upregulated in IL‑1β‑induced chondrocytes, and its knockdown alleviated IL‑1β‑induced chondrocyte injury. SP1 could bind to the ADAMTS5 promoter region to promote its transcription, and SP1 knockdown relieved IL‑1β‑induced chondrocyte injury by reducing ADAMTS5 expression. The SP1/ADAMTS5 axis activated the Wnt/β‑catenin pathway, and the Wnt/β‑catenin pathway agonist, SKL2001, reversed the protective effect of ADAMTS5 knockdown on chondrocyte injury induced by IL‑1β. To the best of our knowledge, the present study was the first to reveal the interaction between SP1 and ADAMTS5 in OA progression and indicated that the SP1/ADAMTS5 axis mediates OA progression by regulating the Wnt/β‑catenin pathway.
骨关节炎（OA）是一种涉及软骨细胞损伤的慢性疾病。ADAMTS5 已被证实介导软骨细胞损伤，从而调节 OA 的进展，但其潜在的分子机制尚不清楚。在本研究中，使用白细胞介素-1β（IL-1β）诱导的软骨细胞来模拟 OA。通过 MTT 测定、EdU 测定和流式细胞术评估细胞增殖和凋亡，通过 Western blot 检测 ADAMTS5、特异性蛋白 1（SP1）、基质相关标志物和 Wnt/β-连环蛋白通路相关标志物的蛋白水平。此外，通过酶联免疫吸附试验测量炎症因子的浓度，并通过检测 SOD 活性和 MDA 水平评估氧化应激。通过逆转录-定量 PCR 测定 ADAMTS5 和 SP1 的 mRNA 表达水平，并通过双荧光素酶报告基因测定和染色质免疫沉淀测定分析 SP1 和 ADAMTS5 之间的相互作用。IL-1β 抑制软骨细胞增殖，但促进凋亡、细胞外基质降解、炎症和氧化应激。ADAMTS5 在 IL-1β 诱导的软骨细胞中上调，其敲低可减轻 IL-1β 诱导的软骨细胞损伤。SP1 可与 ADAMTS5 启动子区域结合，促进其转录，SP1 敲低通过降低 ADAMTS5 表达减轻 IL-1β 诱导的软骨细胞损伤。SP1/ADAMTS5 轴激活 Wnt/β-连环蛋白通路，Wnt/β-连环蛋白通路激动剂 SKL2001 逆转了 ADAMTS5 敲低对 IL-1β 诱导的软骨细胞损伤的保护作用。据我们所知，本研究首次揭示了 SP1 和 ADAMTS5 在 OA 进展中的相互作用，并表明 SP1/ADAMTS5 轴通过调节 Wnt/β-连环蛋白通路介导 OA 的进展。
