Wan Xuan, Xu Pengchao, Zhou Xing, Liu Jiangyuan, Yang Yiwen, Liang Chaoyi, Wang Jinglei, Wang Weixiang, Xu Fengjiao, Wan Xiaoming, Kang Jian, Tong Peijian, Xia Hanting
Department of Orthopedics, Affiliated hospital of Jiangxi University of Chinese Medicine, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi province 330004, China; University Medicine Rostock, University of Rostock, Parkstr. 6, Rostock 18057, Germany.
Zhejiang Provincial Chinese Medicine Hospital (First affiliated hospital of Zhejiang Chinese Medical University), Zhejiang Chinese Medicine University, No. 548, Binwen Road, Binjiang District, Hangzhou City, Zhejiang Province 310053, China.
Phytomedicine. 2025 Jul;142:156764. doi: 10.1016/j.phymed.2025.156764. Epub 2025 Apr 14.
Osteoporosis (OP) represents a systemic disease causing reduced bone mass and fragility fractures. Qigu Capsule (QGC), a traditional Chinese medicine, shows potential in alleviating human OP, but its precise mechanisms remain unclear, limiting clinical application.
The bioactive components of QGC were analyzed using high-performance liquid chromatography (HPLC). An ovariectomy (OVX)-provoked OP rat model was established to evaluate QGC's effects on bone mass, trabecular architecture, and mechanical strength using micro-CT, histological staining, and biomechanical testing. RNA-seq analysis of human OP-derived mesenchymal stem cell (MSC) samples was performed to identify oxidative stress (OxS)- and senescence-associated gene changes. OxS-induced MSC senescence was modeled in vitro using H₂O₂, and QGC's effects on MSC proliferation, migration, and osteogenic differentiation were assessed. Network pharmacology (NP) was deployed to predict the key mechanisms behind the QGC treatment of OP. Further mechanistic studies utilized pharmacological inhibitors and siRNA-mediated gene knockdown to confirm the involvement of critical signaling pathways.
HPLC-MS analysis identified 505 unique bioactive compounds in QGC. In vivo, QGC significantly improved BMD, enhanced trabecular microarchitecture, and restored mechanical properties in OVX rats. ELISA, histological, and immunohistochemical analyses confirmed that QGC primarily enhanced osteoblast activity. RNA-seq analysis of GEO datasets revealed upregulation of senescence and OxS markers (P53, CDKN1A, and INOS) in human OP-derived MSCs. Both in vivo and in vitro QGC alleviated OxS-induced MSC senescence, reduced reactive oxygen species (ROS) levels, suppressed senescence and OxS marker, and promoted MSC proliferation, migration, and osteogenic differentiation. Moreover, NP predicted HIF-1α signaling as critical in QGC's regulation of MSC function during OP. Mechanistic studies demonstrated that QGC activated the HIF-1α/AMPK axis, and inhibition of either HIF-1α or AMPK abolished its therapeutic effects.
QGC mitigates OxS-induced MSC senescence and promotes osteogenesis through the HIF-1α/AMPK axis, highlighting its mechanistic basis in treating OP. These findings show QGC's potential as a therapeutic agent, not only by promoting osteogenesis but also by complementing or serving as an alternative to current OP treatments, offering valuable prospects for enhanced clinical management.
骨质疏松症（OP）是一种导致骨量减少和脆性骨折的全身性疾病。杞骨胶囊（QGC）作为一种传统中药，在缓解人类OP方面显示出潜力，但其确切机制仍不清楚，限制了临床应用。
采用高效液相色谱法（HPLC）分析QGC的生物活性成分。建立卵巢切除（OVX）诱导的OP大鼠模型，使用显微CT、组织学染色和生物力学测试评估QGC对骨量、小梁结构和机械强度的影响。对人OP来源的间充质干细胞（MSC）样本进行RNA测序分析，以确定氧化应激（OxS）和衰老相关基因的变化。使用过氧化氢在体外模拟OxS诱导的MSC衰老，并评估QGC对MSC增殖、迁移和成骨分化的影响。采用网络药理学（NP）预测QGC治疗OP的关键机制。进一步的机制研究利用药理学抑制剂和siRNA介导的基因敲低来证实关键信号通路的参与。
HPLC-MS分析在QGC中鉴定出505种独特的生物活性化合物。在体内，QGC显著提高了OVX大鼠的骨密度，增强了小梁微结构，并恢复了机械性能。ELISA、组织学和免疫组化分析证实，QGC主要增强了成骨细胞活性。对GEO数据集的RNA测序分析显示，人OP来源的MSCs中衰老和OxS标志物（P53、CDKN1A和INOS）上调。在体内和体外，QGC均减轻了OxS诱导的MSC衰老，降低了活性氧（ROS）水平，抑制了衰老和OxS标志物，并促进了MSC增殖、迁移和成骨分化。此外，NP预测HIF-1α信号在QGC调节OP期间MSC功能中起关键作用。机制研究表明，QGC激活了HIF-1α/AMPK轴，抑制HIF-1α或AMPK均可消除其治疗效果。
QGC通过HIF-1α/AMPK轴减轻OxS诱导的MSC衰老并促进成骨，突出了其治疗OP的机制基础。这些发现表明QGC不仅通过促进成骨，还通过补充或替代当前的OP治疗方法，具有作为治疗药物的潜力，为加强临床管理提供了有价值的前景。
