补肾活血胶囊治疗股骨头坏死的分子机制研究
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摘要
目的:基于体内实验、网络药理学和生物信息学探究补肾活血胶囊治疗股骨头坏死(ONFH)的分子机制。方法:体内实验验证补肾活血胶囊治疗ONFH的药效学,利用网络药理学和生物信息学手段确定补肾活血胶囊的活性成分及其治疗ONFH的特异性靶点,分析其信号通路富集程度以探究其可能的分子机制,并进行实验验证。结果:体内动物实验显示补肾活血胶囊能有效地防治激素引起的ONFH大鼠骨丢失情况;通过网络药理学和生物信息学确定了41个补肾活血胶囊治疗ONFH的主要候选靶点;基因富集分析表明,这些靶点与细胞增殖、慢性炎症、脂代谢及血管再生相关信号通路有关;借助体内实验也证实了补肾活血胶囊对上述病理进程的干预机制。结论:补肾活血胶囊治疗ONFH的主要途径为改善脂代谢和调节血管再生;基于网络药理学、生物信息学和实验验证的研究策略为ONFH的发病机制提供新的思路。
Objective: To investigate the molecular mechanism of Bushen Huoxue Capsule in the treatment of osteonecrosis of the femoral head(ONFH) based on in vivo experiments, network pharmacology and bioinformatics analysis.Methods: The pharmacodynamics of Bushen Huoxue Capsule in treating ONFH was determined in vivo. The active components of Bushen Huoxue Capsule and its specific target for treating ONFH were predicted by network pharmacology and bioinformatics analysis. Further signaling pathway enrichment was analyzed to explore its potential molecular mechanism and to be verified experimentally. Results: Animal experiments in vivo showed that Bushen Huoxue Capsule could effectively prevent and treat the bone loss induced by hormone in ONFH rats; Forty-one candidates of Bushen Huoxue Capsule for treating ONFH had been identified by network pharmacology and bioinformatics analysis. These targets were involved in cell proliferation, chronic inflammation, lipid metabolism and vascular regeneration related signal pathway. Conclusion: Bushen Huoxue Capsule has many components and multiple targets in the treatment of osteoarthritis, which is the relevant and multi-level final result with a multiple signal path. The main mechanisms of Bushen Huoxue Capsule in the treatment of ONFH were to improve adipogenic metabolism and regulate vascular regeneration; the strategies based on network pharmacology, bioinformatics and experimental verification provided new ideas for the pathogenesis of ONFH.
Objective: To investigate the molecular mechanism of Bushen Huoxue Capsule in the treatment of osteonecrosis of the femoral head(ONFH) based on in vivo experiments, network pharmacology and bioinformatics analysis.Methods: The pharmacodynamics of Bushen Huoxue Capsule in treating ONFH was determined in vivo. The active components of Bushen Huoxue Capsule and its specific target for treating ONFH were predicted by network pharmacology and bioinformatics analysis. Further signaling pathway enrichment was analyzed to explore its potential molecular mechanism and to be verified experimentally. Results: Animal experiments in vivo showed that Bushen Huoxue Capsule could effectively prevent and treat the bone loss induced by hormone in ONFH rats; Forty-one candidates of Bushen Huoxue Capsule for treating ONFH had been identified by network pharmacology and bioinformatics analysis. These targets were involved in cell proliferation, chronic inflammation, lipid metabolism and vascular regeneration related signal pathway. Conclusion: Bushen Huoxue Capsule has many components and multiple targets in the treatment of osteoarthritis, which is the relevant and multi-level final result with a multiple signal path. The main mechanisms of Bushen Huoxue Capsule in the treatment of ONFH were to improve adipogenic metabolism and regulate vascular regeneration; the strategies based on network pharmacology, bioinformatics and experimental verification provided new ideas for the pathogenesis of ONFH.
引文
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[14]张长青.股骨头坏死保髋治疗指南(2016版).中华老年骨科与康复电子杂志,2016,2(2):65-70
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[16]Shen P,Wong S P,Yong E L.Sensitive and rapid method to quantify icaritin and desmethylicaritin in human serum using gas chromatography-mass spectrometry.J Chromatogr B Analyt Technol Biomed Life Sci,2007,857(1):47-52
[17]Griffith J F,Antonio G E,Kumta S M,et al.Osteonecrosis of hip and knee in patients with severe acute respiratory syndrometreated with steroids.Radiology,2005,235(1):168-175
[18]孙彬,李刚.补肾活血汤治疗股骨头坏死38例观察.实用中医药杂志,2013,29(12):987-988
[19]李凯,贾利利,张振凌.不同煎煮方法黄连阿胶汤对失眠小鼠作用的比较.中国中医基础医学杂志,2013,19(10):1191-1193
[20]Zhang Y L,Yin J H,Ding H,et al.Vitamin K2 Prevents Glucocorticoid-induced Osteonecrosis of the Femoral Head in Rats.Int J Biol Sci,2016,12(4):347-358
[21]Tao S C,Yuan T,Rui B Y,et al.Exosomes derived from human platelet-rich plasma prevent apoptosis induced by glucocorticoidassociated endoplasmic reticulum stress in rat osteonecrosis of the femoral head via the Akt/Bad/Bcl-2 signal pathway.Theranostics,2017,7(3):733-750
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[23]Xue R,Fang Z,Zhang M,et al.TCMID:Traditional Chinese medicine integrative database for herb molecular mechanism analysis.Nucleic Acids Research,2013,41(Database issue):D1089
[24]Xu X,Zhang W,Huang C,et al.A novel chemometric method for the Prediction of Human Oral Bioavailability.International Journal of Molecular Sciences,2012,13(6):6964-6982
[25]Ma C,Wang L,Xie X Q.GPU accelerated chemical similarity calculation for compound library comparison.Journal of Chemical Information&Modeling,2011,51(7):1521-1527
[26]Huang Y A,You Z H,Chen X.A systematic prediction of drug-target interactions using molecular fingerprints and protein sequences.Curr Protein Pept Sci,2018,19(5):468-478
[27]Edgar R,Domrachev M,Lash A E.Gene Expression Omnibus:NCBI gene expression and hybridization array data repository.Nucleic Acids Res,2002,30(1):207-210
[28]Cook H V,Doncheva N T,Szklarczyk D,et al.Viruses.STRING:A Virus-Host Protein-Protein Interaction Database.Viruses,2018,10(10)
[29]Shannon P,Markiel A,Ozier O,et al.Cytoscape:A software environment for integrated models of biomolecular interaction networks.Genome Res,2003,13(11):2498-2504
[30]Huang D W,Sherman B T,Lempicki R A.Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.Nat Protoc,2009,4(1):44-57
[31]Altermann E,Klaenhammer T R.PathwayVoyager:Pathway mapping using the Kyoto Encyclopedia of Genes and Genomes(KEGG)database.BMC Genomics,2005,6:60
[32]赵志强,阎晓霞.中药补肾法改善原发性骨质疏松症临床症状的研究.中国骨质疏松杂志,2018,24(3):371-375
[33]徐宝林.补肾活血胶囊对激素干预大鼠BMSCs增殖及分化的影响.济南:山东中医药大学,2017
[34]Li X,Jin L,Cui Q,et al.Steroid effects on osteogenesis through mesenchymal cell gene expression.Osteoporosis International,2005,16(1):101-108
[35]Mankin H J.Nontraumatic necrosis of bone(osteonecrosis).N Engl J Med,1992,326(22):1473-1479
[36]Huang Z,Cheng C,Cao B,et al.Icariin protects against glucocorticoid-induced osteonecrosis of the femoral head in rats.Cell Physiol Biochem,2018,47(2):694-706
[37]陈娟,邓军,张宇燕,等.丹参素对高脂血症大鼠脂代谢调节机制研究.中国中药杂志,2015,40(2):313-317
[38]李文娜,韩宇东,刘银花,等.杜仲叶绿原酸提取物对脂代谢关键酶活性的影响.中药新药与临床药理,2012,23(1):30-33
[39]王亚玲,崔张新,李方江.丹参与川芎联合用药对冠心病患者脂代谢及血液流变性的影响.时珍国医国药,2009,20(9):2169-2170
[40]张雁儒,张辉,卡卡,等.血管内皮生长因子复合骨形态发生蛋白-2转染骨髓间充质干细胞修复兔股骨头坏死模型.解剖学杂志,2015,38(2):157-161
[41]Xudong L,Quanjun C,Chinghai K,et al.Lovastatin inhibits adipogenic and stimulates osteogenic differentiation by suppressing PPARgamma2 and increasing Cbfa1/Runx2 expression in bone marrow mesenchymal cell cultures.Bone,2003,33(4):652-659
[2]Assoulinedayan Y,Chang C,Greenspan A,et al.Pathogenesis and natural history of osteonecrosis.Seminars in Arthritis&Rheumatism,2002,32(2):94-124
[3]Kaste S C,Pei D,Cheng C,et al.Utility of early screening magnetic resonance imaging for extensive hip osteonecrosis in pediatric patients treated with glucocorticoids.Journal of Clinical Oncology Official Journal of the American Society of Clinical Oncology,2015,33(6):610-615
[4]Duran-Barragan S,Sandoval-Garcia L,Navarrete-Lorenzon M.Additive risk factors resulting in severe osteonecrosis in systemic lupus erythematosus.Arthritis Rheum,2010,62(11):3276
[5]Marston S B,Gillingham K,Bailey R F,et al.Osteonecrosis of the femoral head after solid organ transplantation:a prospective study.J Bone Joint Surg Am,2002,84-A(12):2145-2151
[6]Talamo G,Angtuaco E,Walker R C,et al.Avascular necrosis of femoral and/or humeral heads in multiple myeloma:Results of a prospective study of patients treated with dexamethasonebased regimens and high-dose chemotherapy.J Clin Oncol,2005,23(22):5217-5223
[7]Li X,Jin L,Cui Q,et al.Steroid effects on osteogenesis through mesenchymal cell gene expression.Osteoporos Int,2005,16(1):101-108
[8]Murata M,Kumagai K,Miyata N,et al.Osteonecrosis in stroke-prone spontaneously hypertensive rats:Effect of glucocorticoid.J Orthop Sci,2007,12(3):289-295
[9]Urbaniak J R,Seaber A V,Chen L E.Assessment of ischemia and reperfusion injury.Clin Orthop Relat Res,1997(334):30-36
[10]Chen C,Yang S,Feng Y,et al.Impairment of two types of circulating endothelial progenitor cells in patients with glucocorticoid-induced avascular osteonecrosis of the femoral head.Joint Bone Spine,2013,80(1):70-76
[11]Yeh C H,Chang J K,Wang Y H,et al.Ethanol may suppress Wnt/beta-catenin signaling on human bone marrow stroma cells:A preliminary study.Clin Orthop Relat Res,2008,466(5):1047-1053
[12]Cuadrado M J,Lopez-Pedrera C.Antiphospholipid syndrome.Clin Exp Med,2003,3(3):129-139
[13]Drescher W,Varoga D,Liebs T R,et al.Femoral artery constriction by norepinephrine is enhanced by methylprednisolone in a rat model.J Bone Joint Surg Am,2006,88(Suppl 3):162-166
[14]张长青.股骨头坏死保髋治疗指南(2016版).中华老年骨科与康复电子杂志,2016,2(2):65-70
[15]梁学振,许波,李刚,等.淫羊藿治疗股骨头坏死的分子机制分析.中国实验方剂学杂志,2018,24(14):188-194
[16]Shen P,Wong S P,Yong E L.Sensitive and rapid method to quantify icaritin and desmethylicaritin in human serum using gas chromatography-mass spectrometry.J Chromatogr B Analyt Technol Biomed Life Sci,2007,857(1):47-52
[17]Griffith J F,Antonio G E,Kumta S M,et al.Osteonecrosis of hip and knee in patients with severe acute respiratory syndrometreated with steroids.Radiology,2005,235(1):168-175
[18]孙彬,李刚.补肾活血汤治疗股骨头坏死38例观察.实用中医药杂志,2013,29(12):987-988
[19]李凯,贾利利,张振凌.不同煎煮方法黄连阿胶汤对失眠小鼠作用的比较.中国中医基础医学杂志,2013,19(10):1191-1193
[20]Zhang Y L,Yin J H,Ding H,et al.Vitamin K2 Prevents Glucocorticoid-induced Osteonecrosis of the Femoral Head in Rats.Int J Biol Sci,2016,12(4):347-358
[21]Tao S C,Yuan T,Rui B Y,et al.Exosomes derived from human platelet-rich plasma prevent apoptosis induced by glucocorticoidassociated endoplasmic reticulum stress in rat osteonecrosis of the femoral head via the Akt/Bad/Bcl-2 signal pathway.Theranostics,2017,7(3):733-750
[22]Ru J,Li P,Wang J,et al.TCMSP:A database of systems pharmacology for drug discovery from herbal medicines.Journal of Cheminformatics,2014,6(1):13
[23]Xue R,Fang Z,Zhang M,et al.TCMID:Traditional Chinese medicine integrative database for herb molecular mechanism analysis.Nucleic Acids Research,2013,41(Database issue):D1089
[24]Xu X,Zhang W,Huang C,et al.A novel chemometric method for the Prediction of Human Oral Bioavailability.International Journal of Molecular Sciences,2012,13(6):6964-6982
[25]Ma C,Wang L,Xie X Q.GPU accelerated chemical similarity calculation for compound library comparison.Journal of Chemical Information&Modeling,2011,51(7):1521-1527
[26]Huang Y A,You Z H,Chen X.A systematic prediction of drug-target interactions using molecular fingerprints and protein sequences.Curr Protein Pept Sci,2018,19(5):468-478
[27]Edgar R,Domrachev M,Lash A E.Gene Expression Omnibus:NCBI gene expression and hybridization array data repository.Nucleic Acids Res,2002,30(1):207-210
[28]Cook H V,Doncheva N T,Szklarczyk D,et al.Viruses.STRING:A Virus-Host Protein-Protein Interaction Database.Viruses,2018,10(10)
[29]Shannon P,Markiel A,Ozier O,et al.Cytoscape:A software environment for integrated models of biomolecular interaction networks.Genome Res,2003,13(11):2498-2504
[30]Huang D W,Sherman B T,Lempicki R A.Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.Nat Protoc,2009,4(1):44-57
[31]Altermann E,Klaenhammer T R.PathwayVoyager:Pathway mapping using the Kyoto Encyclopedia of Genes and Genomes(KEGG)database.BMC Genomics,2005,6:60
[32]赵志强,阎晓霞.中药补肾法改善原发性骨质疏松症临床症状的研究.中国骨质疏松杂志,2018,24(3):371-375
[33]徐宝林.补肾活血胶囊对激素干预大鼠BMSCs增殖及分化的影响.济南:山东中医药大学,2017
[34]Li X,Jin L,Cui Q,et al.Steroid effects on osteogenesis through mesenchymal cell gene expression.Osteoporosis International,2005,16(1):101-108
[35]Mankin H J.Nontraumatic necrosis of bone(osteonecrosis).N Engl J Med,1992,326(22):1473-1479
[36]Huang Z,Cheng C,Cao B,et al.Icariin protects against glucocorticoid-induced osteonecrosis of the femoral head in rats.Cell Physiol Biochem,2018,47(2):694-706
[37]陈娟,邓军,张宇燕,等.丹参素对高脂血症大鼠脂代谢调节机制研究.中国中药杂志,2015,40(2):313-317
[38]李文娜,韩宇东,刘银花,等.杜仲叶绿原酸提取物对脂代谢关键酶活性的影响.中药新药与临床药理,2012,23(1):30-33
[39]王亚玲,崔张新,李方江.丹参与川芎联合用药对冠心病患者脂代谢及血液流变性的影响.时珍国医国药,2009,20(9):2169-2170
[40]张雁儒,张辉,卡卡,等.血管内皮生长因子复合骨形态发生蛋白-2转染骨髓间充质干细胞修复兔股骨头坏死模型.解剖学杂志,2015,38(2):157-161
[41]Xudong L,Quanjun C,Chinghai K,et al.Lovastatin inhibits adipogenic and stimulates osteogenic differentiation by suppressing PPARgamma2 and increasing Cbfa1/Runx2 expression in bone marrow mesenchymal cell cultures.Bone,2003,33(4):652-659