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| 苦参黄素调控骨质疏松症破骨细胞分化研究 |
| Study on the regulation of osteoclastogenesis in osteoporosis with kurarinone |
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| DOI：10.3969/j.issn.1006-7108.2025.11.003 |
| 中文关键词: 苦参黄素 破骨细胞 骨髓来源巨噬细胞 RANKL MAPK 骨质疏松症 |
| 英文关键词:kurarinone osteoclast BMDM RANKL MAPK osteoporosis |
| 基金项目:江西省中医药管理局科技计划（2022B194） |
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| 中文摘要: |
| 目的 研究苦参黄素（kurarinone，KE）调控骨质疏松症中破骨细胞分化的作用机制。方法 运用网络药理学和生物信息学分析预测KE调控骨质疏松症中破骨细胞分化影响的主要生物进程和信号通路。以小鼠骨髓来源巨噬细胞（bone marrow-derived macrophage, BMDM）为体外细胞研究对象，使用巨噬细胞集落刺激因子（macrophage colony - stimulating factor, MCSF）和核因子κB受体活化因子配体（receptor activator of nuclear factor kappa-B ligand, RANKL）组成的破骨细胞诱导剂对BMDM进行破骨细胞分化诱导。通过破骨细胞特异性抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase, TRAcP）染色和细胞骨架F-actin环染色检测KE调控破骨细胞分化的作用，使用Western Blot技术对生信分析预测的信号通路进行验证，最后通过荧光染色和流式细胞仪技术检测KE对破骨细胞分化中ROS的影响。结果 网络药理学和生信分析获得KE调控骨质疏松症的蛋白互作网络，核心靶点为TNF、MAPK1、PIK3CA、HSP90AA1和MAPK14等。通路富集发现KE调控破骨细胞分化主要与破骨细胞分化、MAPK信号通路相关，功能富集发现与蛋白激酶活性、MAPK级联调节进程相关。体外细胞实验表明8 μmol/L的KE能显著抑制MAPK信号通路上JNK、P38和ERK的磷酸化水平；抑制破骨细胞分化相关蛋白c-Fos、CTSK、MMP9和NFATc1的表达；减少ROS产生和细胞骨架F-actin环形成；抑制RANKL诱导的BMDM向破骨细胞分化。结论 KE通过激活MAPK信号通路和减少ROS产生抑制破骨细胞分化。 |
| 英文摘要: |
| Objective To investigate the mechanism of action of kurarinone (KE) in regulating osteoclast differentiation in osteoporosis. Methods Network pharmacology and biosignaling analysis were used to predict the major biological processes and signaling pathways involved in the effects of KE-regulated osteoclast differentiation in osteoporosis. Mice bone marrow-derived macrophages (BMDM) were used for in vitro cellular studies. Osteoclastogenesis was induced in BMDM using an osteoclast inducer consisting of macrophage colony-stimulating factor (MCSF) and nuclear factor-κB receptor-activating factor ligand (RANKL). The role of KE in regulating osteoclast differentiation was detected with osteoclast-specific anti-tartrate acid phosphatase (TRAcP) staining and cytoskeletal F-actin ring staining. The signaling pathways predicted by biosynthesis analysis were verified using Western blotting technique. The effect of KE on ROS in osteoclastogenesis was detected using fluorescence staining and flow cytometry technique. Results Network pharmacology and biosignature analyses were performed to obtain the protein interaction network of KE regulation of osteoporosis, with core targets such as TNF, MAPK1, PIK3CA, HSP90AA1, and MAPK14. Pathway enrichment revealed that KE regulation of osteoclastogenesis was mainly associated with osteoclast differentiation and MAPK signaling pathway. Functional enrichment revealed that it was associated with protein kinase activity and MAPK cascade regulatory processes. In vitro cellular experiments showed that 8 μmol/L of KE significantly inhibited the phosphorylation levels of JNK, P38, and ERK in the MAPK signaling pathway, inhibited the expression of osteoclast differentiation-related proteins c-Fos, CTSK, MMP9, and NFATc1, reduced ROS production and cytoskeletal F-actin ring formation, and inhibited RANKL-induced BMDM to osteoclastogenesis. Conclusion KE inhibits osteoclastogenesis by activating the MAPK signaling pathway and by reducing ROS production. |
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