利用网络药理学方法探究天山雪莲药效成分治疗骨肉瘤的分子作用机制  

Exploring the molecular mechanisms of Saussureainvolucrate therapeutic compounds in the treatment of osteosarcoma using network pharmacology

作　　者：陈小雷 孟繁琦 李心乐 张平 CHEN Xiaolei;MENG Fanqi;LI Xinle;ZHANG Ping(Department of Anatomy,School of Basic Medical Sciences,Tianjin Medical University,Tianjin 300070,China;Key Laboratory of Hormones and Development(Ministry of Health),Tianjin Key Laboratory of Metabolic Diseases,Tianjin 300134,China;Tianjin Key Laboratory of Spine and Spinal Cord,Tianjin 300052,China)

机构地区：[1]天津医科大学基础医学院人体解剖学系,天津300070 [2]卫生部激素与发育重点实验室,天津市代谢性疾病重点实验室,天津300134 [3]天津市脊柱脊髓重点实验室,天津300052

出　　处：《天津医科大学学报》2025年第2期111-119,共9页Journal of Tianjin Medical University

摘　　要：目的:探讨天山雪莲药效成分在骨肉瘤治疗中的潜在机制。方法:利用传统中药系统药理学平台(TCMSP)筛选天山雪莲的药效成分和潜在靶点,并通过GEO数据库分析骨肉瘤的差异表达基因。通过韦恩图识别天山雪莲治疗骨肉瘤的潜在靶点,结合蛋白质相互作用(PPI)网络及拓扑学分析筛选核心靶点。随后进行分子对接分析,并通过实时荧光定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法(WB),验证重要药效成分对核心基因表达水平的影响。结果:识别天山雪莲与骨肉瘤的共同靶点67个,确定基质金属蛋白酶2(MMP2)为其中的核心基因。分子对接分析显示,槲皮素与MMP2蛋白具有较强的结合能力(结合能为8.9 kcal/mol)。实验进一步证实槲皮素显著抑制MMP2 mRNA在骨肉瘤细胞系U2OS(7.5μmol/L:t=9.30,P=0.0007;15μmol/L:t=11.97,P=0.0003)及HOS(7.5μmol/L:t=13.09,P=0.0002;15μmol/L:t=21.77,P<0.0001)中的表达水平。此外,槲皮素还显著下调了MMP2蛋白在U2OS(7.5μmol/L:t=9.14,P=0.0008;15μmol/L:t=11.97,P=0.0003)及HOS细胞中的表达(7.5μmol/L:t=20.55,P<0.0001;15μmol/L:t=41.88,P<0.0001)。结论:天山雪莲的关键成分槲皮素可靶向抑制骨肉瘤细胞中MMP2的表达水平,对骨肉瘤具有潜在的治疗价值。Objective:To investigate the potential mechanisms of therapeutic compounds of Saussureainvolucrate(Kar.et Kir.)in the treatment of osteosarcoma.Methods:The therapeutic components and potential targets of Saussureainvolucrata were screened using the Traditional Chinese Medicine Systems Pharmacology(TCMSP)platform,and differentially expressed genes in osteosarcoma were ana-lyzed using the GEO database.Potential targets of Saussureainvolucrate against for osteosarcoma treatment were identified through Venn diagram analysis,and key genes were screened through protein-protein interaction(PPI)network and topological analysis.Molecular docking analysis was subsequently conducted,and the effects of main therapeutic compounds on key gene expression levels were verified using quantitative real-time PCR(qRT-PCR)and Western blotting(WB)assays.Results:A total of 67 common targets between Saussureainvolucrata and osteosarcoma were identified,with matrix metalloproteinase 2(MMP2)recognized as a hub gene.Molecular docking analysis demonstrated a strong binding affinity between quercetin and the MMP2 protein(binding energy of 8.9 kcal/mol).Subsequent experimental results further confirmed that quercetin significantly inhibited the expression levels of MMP2 mRNA in osteosarcoma cell lines U2OS(7.5μmol/L:t=9.30,P=0.0007;15μmol/L:t=11.97,P=0.0003)and HOS(7.5μmol/L:t=13.09,P=0.0002;15μmol/L:t=21.77,P<0.0001).Furthermore,quercetin also significantly reduced protein expression levels of MMP2 in U2OS(7.5μmol/L:t=9.14,P=0.0008;15μmol/L:t=11.97,P=0.0003)and HOS cells(7.5μmol/L:t=20.55,P<0.0001;15μmol/L:t=41.88,P<0.0001).Conclusion:The key component of Saussureainvolucrata,quercetin,can targeted inhibit the expression level of MMP2 in osteosarcoma cells,which has potential therapeutic value for osteosarcoma.

关 键 词：天山雪莲 槲皮素 骨肉瘤 基质金属蛋白酶2 网络药理学 

分 类 号：R932[医药卫生—生药学]