中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (28): 7237-7244.doi: 10.12307/2026.680

• 骨组织构建 bone tissue construction •    下一篇

大黄素甲醚调节骨稳态的功能及分子机制

祁雨心1，2，党一凡1，2，戴黎鸣2，张晓玲1，2   

  1. 1广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心，广西壮族自治区南宁市  530021；2上海交通大学医学院附属新华医院骨科，上海市  200092
  • 收稿日期:2025-06-26 修回日期:2025-10-20 出版日期:2026-10-08 发布日期:2026-02-07
  • 通讯作者: 张晓玲，教授，博士生导师，广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心，广西壮族自治区南宁市 530021；上海交通大学医学院附属新华医院骨科，上海市 200092
  • 作者简介:祁雨心，女，1999年生，汉族，广东省东莞市人，广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心在读硕士，主要从事骨生物学研究。
  • 基金资助:
    国家自然科学基金项目(32370892)，项目负责人：张晓玲

Function and molecular mechanism of physcion in regulating bone homeostasis

Qi Yuxin1, 2, Dang Yifan1, 2, Dai Liming2, Zhang Xiaoling1, 2   

  1. 1Collaborative Innovation Center of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; 2Department of Orthopedics, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Received:2025-06-26 Revised:2025-10-20 Online:2026-10-08 Published:2026-02-07
  • Contact: Zhang Xiaoling, Professor, Doctoral supervisor, Collaborative Innovation Center of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China; Department of Orthopedics, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • About author:Qi Yuxin, MS candidate, Collaborative Innovation Center of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 32370892 (to ZXL)

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摘要：


文题释义：
骨稳态：是指骨骼通过骨的形成、吸收和重塑等动态平衡过程，维持骨组织的结构和功能相对稳定的状态，以保证骨骼的强度、完整性和正常代谢。
大黄素甲醚：是一种天然的蒽醌类化合物，在植物界中分布较为广泛，常见于大黄、枸杞子、巴戟天等具有通脉养骨和补益精气功效的中药材中，是这些中药材发挥药理作用的重要活性成分之一。

背景：已有研究证实大黄素甲醚对骨质疏松症具有保护作用，但其具体作用机制仍不明确。
目的：通过多维度解析大黄素甲醚对AKT信号通路的调控作用，揭示其对破骨细胞诱导分化和成骨功能诱导分化调节的分子机制。
方法：①体外培养RAW264.7细胞和C3H10T1/2细胞，分别加入0，10，20，30，40，50，60 µmol/L大黄素甲醚处理两种细胞，CCK8法检测大黄素甲醚的细胞毒性。②诱导RAW264.7细胞破骨分化和C3H10T1/2细胞成骨分化，分别给予不同浓度(0，20，40 µmol/L)的大黄素甲醚处理，通过qPCR、Western Blot、碱性磷酸酶染色分别检测分化能力。③基于网络药理学分析大黄素甲醚对破骨细胞分化功能的调控及相关的信号通路，并针对目标靶点蛋白进行分子对接。④Western Blot检测验证大黄素甲醚调节下游靶信号通路AKT轴的AKT磷酸化水平。
结果与结论：①大黄素甲醚在0-60 µmol/L浓度下，各组细胞存活率皆大于90%，无明显细胞毒性。②大黄素甲醚显著抑制破骨细胞分化相关基因表达，Acp5、CTSK、DC-STAMP、Nfatc1呈下调趋势，但对成骨细胞分化相关基因COL1A1、Runx2、OSX表达及碱性磷酸酶染色强度无显著影响。③据网络药理学及分子对接推断，大黄素甲醚影响破骨细胞分化并调控PI3K-AKT通路，与AKT1靶点结合能为-10.72 kJ/mol，
具有较强结合活性。④在破骨分化过程中，RAW264.7细胞的p-AKT/AKT比值升高(n=3，P=0.006 3)，而大黄素甲醚使该比值降低。说明大黄素甲醚通过调节AKT信号通路抑制破骨细胞分化，从而调控骨平衡。 
https://orcid.org/0009-0000-9028-8260(祁雨心)；https://orcid.org/0000-0002-0134-1347(张晓玲)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 网络药理学, 大黄素甲醚, 骨稳态, 破骨细胞, 成骨细胞

Abstract: BACKGROUND: Although physcion has been shown to have protective effects against osteoporosis, the exact mechanism is not fully understood.
OBJECTIVE: Through multidimensional analysis of the regulatory effect of physcion on the AKT signaling pathway, the molecular mechanism of its regulation on osteoclast induced differentiation and osteogenic function induced differentiation is revealed.
METHODS: (1) RAW264.7 cells and C3H10T1/2 cells were cultured in vitro and subsequently exposed to 0, 10, 20, 30, 40, 50, and 60 µmol/L physcion, respectively. The cytotoxicity of physcion was detected by cell counting kit-8 assay. (2) RAW264.7 cells and C3H10T1/2 cells were treated with different concentrations (0, 20, and 40 µmol/L) of physcion during osteoclast and osteogenic differentiation, respectively. The differentiation ability was detected by quantitative polymerase chain reaction, western blot assay, and alkaline phosphatase staining, respectively. (3) Based on network pharmacology, the regulation of osteoclast differentiation by physcion and related pathways was analyzed, and molecular docking was performed for target proteins. (4) Western blot assay was used to verify the regulatory effect of physcion on the phosphorylation of AKT in the AKT axis—a downstream pathway.
RESULTS AND CONCLUSION: (1) Exposure to concentrations of 0-60 µmol/L of physcion resulted in cell survival rates of over 90% in all groups, with no significant cytotoxicity observed. (2) Physcion significantly inhibited the expression of genes related to osteoclast differentiation, with Acp5, CTSK, DC-STAMP, and Nfatc1 showing a downregulation trend. However, it has no significant effect on the expression of genes related to osteoblast differentiation, COL1A1, Runx2, OSX, and alkaline phosphatase staining intensity. (3) According to network pharmacology and molecular docking, physcion affects osteoclast differentiation and regulates the PI3K-AKT pathway, with a binding energy of -10.72 kJ/mol to the AKT1 target and strong binding activity. (4) During osteoclast differentiation, the p-AKT/AKT ratio of RAW264.7 cells was increased (n=3, P=0.006 3), while physcion decreased this ratio. These findings demonstrate that physcion inhibits osteoclast differentiation by regulating the AKT signaling pathway, thereby regulating bone homeostasis. 

Key words: network pharmacology, physcion, bone homeostasis, osteoclast, osteoblast

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