AM1241 inhibits chondrocyte inflammation and ECM degradation through the Nrf2/HO-1 and NF-κB pathways and alleviates osteoarthritis in mice.

IF 6 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Medicine Pub Date : 2025-01-10 DOI:10.1186/s10020-024-01012-5
Zhuan Zou, Songmu Pan, Changzheng Sun, Jiyong Wei, Yi Xu, Kaizhen Xiao, Jinmin Zhao, Ronghe Gu
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Abstract

Background: This study aimed to investigate the impact of AM1241 on lipopolysaccharide (LPS)-induced chondrocyte inflammation in mice and its potential mechanism for improving osteoarthritis (OA).

Methods: The OA mice model was established employing the refined Hulth method. The impact of different concentrations of AM1241 on mice chondrocyte activity was detected using CCK-8. Changes in the levels of LPS-induced inflammatory factors and cartilage extracellular matrix (ECM) degradation in chondrocytes were determined by western blot, RT-qPCR, ELISA, and immunofluorescence assays, respectively. The specific action modes and binding sites of AM1241 with NEMO/IκB kinases (IKKs) in the NF-κB pathway and Keap1 protein in the Nrf2 pathway were predicted via molecular docking and molecular dynamics simulation, and the NF-κB and Nrf2 pathways were detected using western blot and immunofluorescence. In vivo, the impact of AM1241 on OA mice was analyzed through safranin-fast green staining, IHC staining, Mankin score, and microCT.

Results: AM1241 inhibited the levels of LPS-induced transforming growth factor-β (TGF-β1), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), matrix metalloproteinase-13 (MMP-13), and a disintegrin and metalloproteinase with thrombospondin motif 5 (ADAMTS-5) and diminished the degradation of type II collagen and Aggrecan. For the mechanism, AM1241 regulated the NF-kB and Nrf2/HO-1 signaling pathways by binding to NEMO/IKKβ and Keap1 target proteins and suppressed the activation of the NF-κB signaling pathway by activating the Nrf2 in chondrocytes. In vivo, AM1241 inhibited bone anabolism, mitigated articular cartilage hyperplasia and wear, and reduced the Mankin score in mice, thereby hindering the development of OA.

Conclusion: AM1241 inhibited activation of the NF-κB signaling pathway via activating Nrf2. It suppressed the expression of inflammation factors and the degradation of ECM in vitro, and improved OA in mice in vivo, suggesting its potential as an effective drug candidate for the treatment of OA. The remarkable efficacy of AM1241 in alleviating murine OA positions it as a potential therapeutic strategy in the clinical management of OA diseases.

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背景:本研究旨在探讨AM1241对脂多糖(LPS)诱导的小鼠软骨细胞炎症的影响及其改善骨关节炎(OA)的潜在机制:方法:采用改良 Hulth 法建立 OA 小鼠模型。方法:采用改良 Hulth 法建立 OA 小鼠模型,使用 CCK-8 检测不同浓度的 AM1241 对小鼠软骨细胞活性的影响。分别通过 Western blot、RT-qPCR、ELISA 和免疫荧光检测 LPS 诱导的炎症因子和软骨细胞外基质(ECM)降解水平的变化。通过分子对接和分子动力学模拟预测了AM1241与NF-κB通路中的NEMO/IκB激酶(IKKs)和Nrf2通路中的Keap1蛋白的特异性作用模式和结合位点,并利用Western印迹和免疫荧光检测了NF-κB和Nrf2通路。在体内,通过黄绿素-快绿染色、IHC染色、Mankin评分和microCT分析了AM1241对OA小鼠的影响:结果:AM1241抑制了LPS诱导的转化生长因子-β(TGF-β1)、肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)、基质金属蛋白酶-13(MMP-13)和具有凝血酶原基序5的崩解酶和金属蛋白酶(ADAMTS-5)的水平,并减少了II型胶原和Aggrecan的降解。在机制方面,AM1241通过与NEMO/IKKβ和Keap1靶蛋白结合来调节NF-kB和Nrf2/HO-1信号通路,并通过激活软骨细胞中的Nrf2来抑制NF-κB信号通路的激活。在体内,AM1241可抑制骨合成代谢,减轻关节软骨增生和磨损,降低小鼠的Mankin评分,从而阻碍OA的发展:结论:AM1241通过激活Nrf2抑制了NF-κB信号通路的激活。结论:AM1241 通过激活 Nrf2 抑制了 NF-κB 信号通路的激活,在体外抑制了炎症因子的表达和 ECM 的降解,在体内改善了小鼠的 OA,表明它有可能成为治疗 OA 的有效候选药物。AM1241 在缓解小鼠 OA 方面的显著疗效使其成为临床治疗 OA 疾病的一种潜在治疗策略。
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来源期刊
Molecular Medicine
Molecular Medicine 医学-生化与分子生物学
CiteScore
8.60
自引率
0.00%
发文量
137
审稿时长
1 months
期刊介绍: Molecular Medicine is an open access journal that focuses on publishing recent findings related to disease pathogenesis at the molecular or physiological level. These insights can potentially contribute to the development of specific tools for disease diagnosis, treatment, or prevention. The journal considers manuscripts that present material pertinent to the genetic, molecular, or cellular underpinnings of critical physiological or disease processes. Submissions to Molecular Medicine are expected to elucidate the broader implications of the research findings for human disease and medicine in a manner that is accessible to a wide audience.
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