Specnuezhenide attenuates rheumatoid arthritis bone destruction via dual regulation of osteoclast-osteoblast balance through KEAP1/NRF2 signaling

IF 8.3 1区医学 Q1 CHEMISTRY, MEDICINAL

Phytomedicine Pub Date : 2025-09-23 DOI:10.1016/j.phymed.2025.157293

Huan Yao , Qingman He , Jiayue Yang , Li Xiang , Yuanyuan Luo , Zhimin Chen , Xueping Li

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Abstract

Background

Rheumatoid arthritis (RA)-associated bone erosion is driven by osteoclast-osteoblast imbalance and oxidative stress. Specnuezhenide (SPN), a bioactive compound from Ligustrum lucidum, exhibits antioxidant properties, but its role in RA bone loss remains undefined.

Purpose

To explore the potential mechanisms and therapeutic targets of SPN in treating RA, with a focus on its regulation of bone remodeling and oxidative stress pathways.

Study Design

Collagen-induced arthritis (CIA) mice were treated with SPN (50/100/200 mg/kg) for 28 days. Joint inflammation, bone erosion, osteoclast/osteoblast markers, and oxidative stress pathways were evaluated. In vitro, SPN’s effects on osteoclastogenesis/osteoblastogenesis were assessed in inflammatory microenvironments. Molecular docking, dynamics simulations, and KEAP1 knockdown experiments were used to validate mechanisms.

Methods

In vivo, CIA was induced in DBA/1 mice, followed by SPN treatment. Paw swelling, arthritis index (AI), micro-CT for bone erosion, H&E and TRAP staining for histological analysis, Western blotting for marker proteins, and immunofluorescence for NRF2 localization were assessed. In vitro, osteoclast/osteoblast differentiation from bone marrow cells under inflammatory conditions (LPS/nigericin) was evaluated. Mechanistic investigations were performed via molecular docking, molecular dynamics simulations, cellular thermal shift assay, surface plasmon resonance, and adenovirus-mediated KEAP1 knockdown.

Results

In vivo, SPN reduced paw swelling, AI, and bone erosion in CIA mice. It downregulated osteoclast markers (TRAP, NFATC1) and upregulated osteoblast markers (RUNX2, Col1), suppressed ROS, and activated NRF2/SOD1 pathways. In vitro, SPN inhibited osteoclastogenesis and promoted osteoblast mineralization in inflammatory microenvironments, with ROS scavenging dependent on NRF2 activation. Mechanistically, SPN bound to KEAP1, promoting NRF2 nuclear translocation.

Conclusion

SPN attenuates RA bone destruction by restoring osteoclast-osteoblast balance through precise targeting of the KEAP1, thereby modulating the KEAP1/NRF2/ROS axis. This mechanism-specific phytotherapeutic strategy provides a novel approach against RA-related osteolysis distinct from broad-spectrum antioxidants.

Abstract Image

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Specnuezhenide通过KEAP1/NRF2信号通路双重调节破骨细胞-成骨细胞平衡，减轻类风湿关节炎骨破坏

类风湿性关节炎（RA）相关的骨侵蚀是由破骨细胞-成骨细胞失衡和氧化应激驱动的。Specnuezhenide （SPN）是一种来自女贞子的生物活性化合物，具有抗氧化特性，但其在RA骨质流失中的作用尚不清楚。目的探讨SPN治疗RA的潜在机制和治疗靶点，重点探讨其对骨重塑和氧化应激通路的调控作用。实验设计采用SPN （50/100/200 mg/kg）治疗胶原性关节炎（CIA）小鼠28 d。评估关节炎症、骨侵蚀、破骨细胞/成骨细胞标志物和氧化应激途径。体外，在炎症微环境中评估SPN对破骨细胞/成骨细胞发生的影响。通过分子对接、动力学模拟和KEAP1敲低实验来验证其机制。方法在DBA/1小鼠体内诱导CIA，再加SPN处理。评估足跖肿胀、关节炎指数（AI）、骨侵蚀的micro-CT、H&；E和TRAP染色进行组织学分析、Western blotting检测标记蛋白、免疫荧光检测NRF2定位。体外，在炎症条件下（LPS/尼日利亚菌素）评估骨髓细胞的破骨细胞/成骨细胞分化。通过分子对接、分子动力学模拟、细胞热移测定、表面等离子体共振和腺病毒介导的KEAP1敲低进行机制研究。结果在体内，SPN可减轻CIA小鼠足跖肿胀、AI和骨侵蚀。它下调破骨细胞标志物（TRAP、NFATC1）和上调成骨细胞标志物（RUNX2、Col1），抑制ROS，激活NRF2/SOD1通路。在体外，SPN抑制炎症微环境中的破骨细胞生成，促进成骨细胞矿化，ROS清除依赖于NRF2的激活。机制上，SPN与KEAP1结合，促进NRF2核易位。结论spn通过精确靶向KEAP1，调节KEAP1/NRF2/ROS轴，恢复破骨细胞-成骨细胞平衡，从而减轻RA骨破坏。这种机制特异性的植物治疗策略为抗ra相关的骨溶解提供了一种不同于广谱抗氧化剂的新方法。

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来源期刊

Phytomedicine 医学-药学

CiteScore

10.30

自引率

5.10%

发文量

670

审稿时长

91 days

期刊介绍： Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.