Zhou Yi, Li Shaoshuo, Hong Bowen, Wang Zihan, Shao Yang, Wu Mao, Wang Jianwei
Graduate School, Nanjing University of Chinese Medicine, Nanjing, China.
Wuxi Affiliated Hospital of Nanjing University of Chinese Medicine, Wuxi, China.
Front Pharmacol. 2025 Apr 28;16:1551257. doi: 10.3389/fphar.2025.1551257. eCollection 2025.
Osteoporosis (OP) represents a widespread bone remodeling disorder within the domain of orthopedics, markedly compromising the quality of life in the elderly population. The need to develop more efficient therapeutic approaches to attenuate bone resorption by suppressing the excessive activation of osteoclasts (OCs) remains urgent. The plant flavonoid Isorhamnetin (Iso), recognized for its potent antioxidant properties, has been the subject of extensive research regarding its potential in treating bone-related conditions.
This study adopts a comprehensive methodology to evaluate Iso's impact on bone metabolism and its therapeutic possibilities for treating OP. By integrating network pharmacology, molecular dynamics simulations, and surface plasmon resonance (SPR), we performed phenotypic analyses to systematically evaluate the inhibitory effect of Iso on OC differentiation. The mechanisms behind Iso's inhibition of OC differentiation were further elucidated. testing was also performed to substantiate the therapeutic effects of Iso in an OP animal model.
At low concentrations, Iso showed no cytotoxicity and did not interfere with cell proliferation in RAW 264.7 cells. Iso effectively inhibited RANKL-induced osteoclast differentiation in these cells, while downregulating related genes levels (, k, , ). Molecular dynamics simulations and surface plasmon resonance confirmed Iso's dual binding to both RANKL and RANK. KEGG pathway enrichment analysis results indicated that Iso modulates the MAPK, NF-κB/PI3K-AKT, and calcium signaling pathways. Western blot analysis revealed that Iso treatment targeting the RANKL/RANK binding pathway significantly downregulated phosphorylation levels of JNK, P38, AKT, and p65. Concurrently, Iso stimulation markedly increased IκBα expression, thereby rescuing its degradation. Furthermore, Iso demonstrated a robust inhibitory effect on reactive oxygen species levels . Furthermore, in OVX mice, Iso treatment increased bone density, modulated serum bone metabolism markers, and downregulated transcriptional levels of OC marker genes.
Iso exhibits therapeutic potential for OP by selectively targeting and disrupting the RANKL-RANK interaction. This intervention modulates the expression of intracellular transcription factors and multiple signaling pathways, thereby inhibiting the maturation of OCs. Through mitigating OC-mediated bone loss, Iso holds significant promise as a potent therapeutic agent for OP.
骨质疏松症（OP）是骨科领域中一种广泛存在的骨重塑疾病，严重影响老年人群的生活质量。开发更有效的治疗方法以通过抑制破骨细胞（OC）的过度活化来减少骨吸收的需求仍然迫切。植物黄酮异鼠李素（Iso）因其强大的抗氧化特性而受到关注，关于其在治疗骨相关疾病方面的潜力已进行了广泛研究。
本研究采用综合方法评估Iso对骨代谢的影响及其治疗OP的可能性。通过整合网络药理学、分子动力学模拟和表面等离子体共振（SPR），我们进行了表型分析，以系统评估Iso对OC分化的抑制作用。进一步阐明了Iso抑制OC分化的机制。还进行了测试以证实Iso在OP动物模型中的治疗效果。
在低浓度下，Iso对RAW 264.7细胞无细胞毒性且不干扰细胞增殖。Iso有效抑制这些细胞中RANKL诱导的破骨细胞分化，同时下调相关基因水平（，k，，）。分子动力学模拟和表面等离子体共振证实Iso与RANKL和RANK均具有双重结合。KEGG通路富集分析结果表明，Iso调节MAPK、NF-κB/PI3K-AKT和钙信号通路。蛋白质印迹分析显示，针对RANKL/RANK结合途径的Iso处理显著下调JNK、P38、AKT和p65的磷酸化水平。同时，Iso刺激显著增加IκBα表达，从而挽救其降解。此外，Iso对活性氧水平具有强大的抑制作用。此外，在去卵巢小鼠中，Iso治疗增加了骨密度，调节了血清骨代谢标志物，并下调了OC标志物基因的转录水平。
Iso通过选择性靶向和破坏RANKL-RANK相互作用展现出治疗OP的潜力。这种干预调节细胞内转录因子的表达和多种信号通路，从而抑制OC的成熟。通过减轻OC介导的骨质流失，Iso作为一种有效的OP治疗药物具有巨大的前景。
