中药新药与临床药理2025，Vol.36Issue(9)：1471-1482,12.DOI:10.19378/j.issn.1003-9783.2025.09.007
川续断皂苷Ⅵ调控骨相关细胞介导的巨噬细胞M1型极化改善激素性股骨头坏死的机制
Mechanistic Study on Asperosaponin Ⅵ Regulating Bone-Related Cell-Mediated Macrophage M1 Polarization to Improve Steroid-Induced Osteonecrosis of the Femoral Head
摘要
Abstract
Objective To investigate the mechanism of Asperosaponin Ⅵ(ASD)in improving bone-related cell damage in steroid-induced osteonecrosis of the femoral head(SONFH)based on bioinformatics,molecular docking technology and experimental validation.Methods Bioinformatics and network pharmacology study:Differentially expressed genes(DEGs)of SONFH were screened from the GEO database for enrichment analysis to explore disease-related mechanisms.Predicted ASD targets were intersected with disease DEGs to construct a protein-protein interaction(PPI)network.Key therapeutic targets were identified for enrichment analysis and validated via molecular docking to elucidate ASD's potential molecular mechanisms in SONFH treatment.Clinical study:Femoral head specimens from SONFH patients were collected,and inflammatory pathology in necrotic areas was observed using HE staining and immunofluorescence.In vitro experiments:Dexamethasone(Dex)-treated MC3T3-E1 and MLOY4 cells were used to establish an in vitro SONFH model.Results were validated using CCK-8,Calcein/PI cell viability and toxicity assays,qPCR with a co-culture system.Results Bioinformatics and network pharmacology:PPI analysis identified seven key targets(TNF-α,IL-1β,IL-6,etc.),with enrichment analysis revealing strong associations with inflammatory responses and TNF,NF-κB signaling pathways.Clinical study:HE staining showed increased inflammatory cell infiltration in necrotic versus non-necrotic groups(P＜0.05).Immunofluorescence demonstrated elevated iNOS and CD86(P＜0.01)but reduced CD206(P＜0.05)in necrotic areas.In vitro experiments:First-round CCK-8 assays confirmed Dex suppressed MC3T3-E1 and MLOY4 proliferation(P＜0.01,P＜0.05).qPCR showed damaged bone cells upregulated RAW264.7 mRNA expression of TNF-α,IL-1β,IL-6,iNOS,and CD86(P＜0.05,P＜0.01,P＜0.001)while downregulating CD206 mRNA(P＜0.05,P＜0.001).M1-polarized RAW264.7 reduced bone cell proliferation(P＜0.05,P＜0.001)and viability(P＜0.05).Second-round CCK-8 and Calcein/PI assays revealed ASD+Dex improved bone cell proliferation(P＜0.01,P＜0.05)and viability(P＜0.05)versus Dex alone.qPCR showed recovered bone cells downregulated RAW 264.7 mRNA of TNF-α、IL-1β,IL-6,iNOS and CD86(P＜0.05,P＜0.01,P＜0.001)and upregulated CD206 mRNA(P＜0.05)in macrophages.Conclusion ASD alleviates SONFH progression,potentially by inhibiting M1 polarization of macrophages induced by damaged bone-related cells.关键词
激素性股骨头坏死/川续断皂苷Ⅵ/巨噬细胞/炎症反应/生物信息学/网络药理学/实验验证Key words
Steroid-induced osteonecrosis of the femoral head(SONFH)/Asperosaponin Ⅵ/macrophages/inflammatory response/bioinformatics/network pharmacology/experimental volidation分类
医药卫生引用本文复制引用
赵洋,杜希望,韦标方..川续断皂苷Ⅵ调控骨相关细胞介导的巨噬细胞M1型极化改善激素性股骨头坏死的机制[J].中药新药与临床药理,2025,36(9):1471-1482,12.基金项目
2024年度国家级中医药继续教育项目(T20241520005). （T20241520005）
