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目的：探究壮骨止痛方治疗绝经后骨质疏松症的潜在作用靶点与机制。方法：利用STRING数据库建立基于BATMAN-TCM数据库筛选出的药物作用靶点与GeneCards、TTD和OMIM数据库中PMOP疾病靶点交集的PPI网络，并借助Cytoscape软件，将该网络可视化处理，得到“药物-活性成分-靶点-疾病”网络，同时，分子对接验证核心活性成分与关键靶点间的结合能力。实验建立绝经后骨质疏松的动物模型，采用mirco-CT和HE染色技术研究壮骨止痛方对绝经后骨质疏松症的治疗作用，并通过RT-PCR技术评估关键靶点mRNA的表达情况。结果：获得壮骨止痛方活性成分与疾病共同靶点69个。其中关键靶点为PPARG、PTGS2、ESR1等。药物-活性成分-靶点-疾病网络揭示核心活性成分为豆甾醇、异补骨脂素、熊果酸、齐墩果酸等。分子对接结果显示两者有较强结合能力。GO功能分析得到主要生物功能为对激素的反应等。KEGG富集主要相关通路为癌症的发病途径、雌激素信号通路等。CT扫描、HE染色可得壮骨止痛方能有效缓解绝经后骨质疏松症，RT-PCR显示壮骨止痛方组PTGS2mRNA表达显著升高，PPARG表达明显降低（P<0.05）。结论：壮骨止痛方通过多成分、多靶点、多通路治疗PMOP，其成分主要为豆甾醇、异补骨脂素、熊果酸、齐墩果酸，并通过代谢途径、雌激素信号通路等作用于人体INS、AKT1、PPARG、PTGS2、ESR1发挥疗效。
Abstract：Objective: To explore the potential target and mechanism of Zhuanggu Zhitong formula in the treatment of postmenopausal osteoporosis. Methods: The PPI network of drug action targets was screened from the BATMAN-TCM database and PMOP disease targets were obtained from GeneCards, TTD, and OMIM databases. This network was then established using the STRING database and visualized with Cytoscape software to create a ′drug-active ingredient-target-disease′ network. Additionally, molecular docking was employed to confirm the binding ability between core active components and key targets. Molecular docking technique was used to verify the binding ability of core active components to key targets. The animal model of postmenopausal osteoporosis was established. Mirco-CT and HE staining were used to explore the effect of Zhuanggu Zhitong formula in the treatment of postmenopausal osteoporosis. RT-PCR was used to evaluate the expression of mRNA.Results: 69 active ingredient targets and disease common targets of Zhuanggu Zhitong formula were obtained. The key targets of PPI analysis were PPARG, PTGS2, ESR1 and so on. Drug-active ingredient-target-disease network revealed that the core active components were stigmasterol, isopsoralen, ursolic acid, oleanolic acid and so on. The results of molecular docking showed that they had strong binding ability. GO function analysis showed that the main biological function was the response to hormones. The main pathways related to KEGG enrichment were the pathogenesis of cancer, estrogen signal pathway and so on. CT scanning and HE staining can effectively relieve postmenopausal osteoporosis. RT-PCR showed that the expression of PTGS2mRNA was significantly increased and the expression of PPARG was significantly decreased in Zhuanggu Zhitong group.Conclusion: Zhuanggu Zhitong formula can treat PMOP with multi-components, multi-targets and multi-pathways. The main components of Zhuanggu Zhitong formula are stigmasterol, Angelicin, ursolic acid and oleanolic acid. Zhuanggu Zhitong formula acts on human INS, AKT1, PPARG, PTGS2and ESR1 through metabolic pathway and estrogen signal pathway.
1 Deng YX,He WG,Cai HJ,et al.Analysis and validation of Hub genes in blood monocytes of postmenopausal osteoporosis patients[J].Front Endocrinol(Lausanne),2021,12:815245.
2 Arceo-Mendoza RM,Camacho PM.Postmenopausal osteoporosis:Latest guidelines[J].Endocrinol Metab Clin North Am,2021,50(2):167-178.
3 Walker MD,Shane E.Postmenopausal osteoporosis[J].N Engl J Med,2023,389(21):1979-1991.
4 Luo MH,Zhao JL,Xu NJ,et al.Comparative efficacy of Xianling Gubao Capsules in improving bone mineral density in postmenopausal osteoporosis:A network meta-analysis[J].Front Endocrinol(Lausanne),2022,13:839885.
5 Zeng Q,Li N,Wang Q,et al.The prevalence of osteoporosis in China,a nationwide,multicenter DXA survey[J].J Bone Miner Res,2019,34(10):1789-1797.
6 Rozenberg S,Al-Daghri N,Aubertin-Leheudre M,et al.Is there a role for menopausal hormone therapy in the management of postmenopausal osteoporosis?[J].Osteoporos Int,2020,31(12):2271-2286.
7曾英，章文娟，李伟娟等.基于雌激素信号通路的壮骨止痛胶囊治法组方机理探讨[J].时珍国医国药，2017,28(4):868-871.Zeng Y,Zang WJ,Li WJ,et al.Exploration of the mechanism of the therapeutic formula of Strong Bone Pain Relief Capsules based on estrogen signaling pathway[J].Lishizhen Med Mater Med Res,2017,28(4):868-871.
8李劲平，曾英，雷晓明等.壮骨止痛胶囊抗实验性绝经后骨质疏松作用的三维组学分析[J].中国中药杂志，2014,39(16):3203-3207.Li JP,Zeng Y,Lei XM,et al.Tri-dimensional omics analysis on effect of zhuanggu zhitong capsule against experimental postmenopausal osteoporosis[J].Chin JChin Materia Med,2014,39(16):3203-3207.
9蔡昕瑶，陈瑶，陈诗淇等.壮骨止痛方通过RANKL/RANK信号通路抑制骨吸收的机制研究[J].中国骨质疏松杂志，2023,29(9):1333-1339.Cai XY,Chen Y,Chen SQ,et al.Study of the mechanism of inhibition of bone resorption with the strong bone and analgesic prescription through RANKL/RANK signaling pathway[J].Chin J Osteoporosis,2023,29(9):1333-1339.
10 Liu Z,Guo F,Wang Y,et al.BATMAN-TCM:A bioinformatics analysis tool for molecular mech ANism of traditional Chinese medicine[J].Sci Rep,2016,6:21146.
11 Stelzer G,Rosen N,Plaschkes I,et al.The Genecards suite:From gene data mining to disease genome sequence analyses[J].Curr Protoc Bioinformatics,2016,54:1.30.1-1.30.33.
12 Zhou Y,Zhang Y,Lian X,et al.Therapeutic target database update 2022:Facilitating drug discovery with enriched comparative data of targeted agents[J].Nucleic Acids Res,2022,50(D1):D1398-D1407.
13 Amberger JS,Hamosh A.Searching online mendelian inheritance in man(OMIM):A knowledgebase of human genes and genetic phenotypes[J].Curr Protoc Bioinformatics,2017,58:1.2.1-1.2.12.
14 Bardou P,Mariette J,EscudiéF,et al.jvenn:an interactive Venn diagram viewer[J].BMC Bioinformatics,2014,15(1):293.
15 Szklarczyk D,Kirsch R,Koutrouli M,et al.The STRING database in 2023:Protein-protein association networks and functional enrichment analyses for any sequenced genome of interest[J].Nucleic Acids Res,2023,51(D1):D638-D646.
16 Shannon P,Markiel A,Ozier O,et al.Cytoscape:A software environment for integrated models of biomolecular interaction networks[J].Genome Res,2003,13(11):2498-2504.
17 Zhou Y,Zhou B,Pache L,et al.Metascape provides a biologist-oriented resource for the analysis of systemslevel datasets[J].Nat Commun,2019,10(1):1523.
18 Tang D,Chen M,Huang X,et al.SRplot:A free online platform for data visualization and graphing[J].PLo S One,2023,18(11):e0294236.
19 Kim S,Chen J,Cheng T,et al.Pub Chem in 2021:New data content and improved web interfaces[J].Nucleic Acids Res,2021,49(D1):D1388-D1395.
20 Berman HM,Westbrook J,Feng Z,et al.The protein data bank[J].Nucleic Acids Res,2000,28(1):235-242.
21 Trott O,Olson AJ.Auto Dock Vina:Improving the speed and accuracy of docking with a new scoring function,efficient optimization,and multithreading[J].JComput Chem,2010,31(2):455-461.
22陈沙，王桂云，李荣慧等.雌性大鼠手术去势骨质疏松症模型建立及评价[J].湖南中医药大学学报，2020,40(11):1315-1319.Chen S,Wang GY,Li RH,et al.Establishment of osteoporosis model by surgical castration in female rats and its evaluation[J].J Hunan Univ Chin Med,2020,40(11):1315-1319.
23甘国兴，刘毓，李劲平等.壮骨止痛方对去卵巢骨质疏松大鼠CGRP及β2-AR的影响[J].中国医药导报，2018,15(07):9-11,15,181.Gan XG,Liu Y,Li JP,et al.Effect of Zhuanggu Zhitong prescription on CGRP and β2-AR in ovariectomized osteoporosis rats[J].Chin Med Herald,2018,15(7):9-11,15,181.
24杨珊珊，陈红琼，陈瑶等.壮骨止痛胶囊对去卵巢大鼠体质量及脂/骨代谢的影响[J].湖南中医药大学学报，2022,42(1):18-24.Yang SS,Chen HQ,Chen Y,et al.Effect of Zhuanggu Zhitong Capsule on body weight and lipid/bonemetabolism in ovariectomized rats[J].J Hunan Univ Chin Med,2022,42(1):18-24.
25 Ming Y,Jiachen L,Tao G,et al.Exploration of the mechanism of Tripterygium Wilfordii in the treatment of myocardial fibrosis based on network pharmacology and molecular docking[J].Curr Comput Aided Drug Des,2023,19(1):68-79.
26袁玲丹，宋利格.《原发性骨质疏松症诊疗指南（2022版）》解读[J].同济大学学报（医学版），2023,44(6):777-784.Yuan LD,Song LG.Interpretation of the Guidelines for the Diagnosis and Treatment of Primary Osteoporosis(2022)[J].J Tongji Univ(Medical Science).2023,44(6):777-784.
27雷晓明，田松.壮骨止痛胶囊含药血清对大鼠成骨样细胞增殖分化影响的实验研究[J].天津中医药，2004,(3):237-239.Lei XM,Tian S,et al.Effect of Zhuanggu Zhitong Capsule on the proliferation and differentiation in rat of osteoblast[J].Tianjin J Tradit Chin Med,2004,2004(3):237-239.
28 Sampath S,Kotikalapudi N,Venkatesan V.A novel therapeutic combination of mesenchymal stem cells and stigmasterol to attenuate osteoarthritis in rodent model system-a proof of concept study[J].Stem Cell Investig,2021,8:5.
29 Mo Z,Xu P,Li H.Stigmasterol alleviates interleukin-1beta-induced chondrocyte injury by down-regulatingsterol regulatory element binding transcription factor 2 to regulateferroptosis[J].Bioengineered,2021,12(2):9332-9340.
30 Li H,Wang C,Jin Y,et al.Anti-Postmenopausal osteoporosis effects of Isopsoralen:A bioinformatics-integrated experimental study[J].Phytother Res,2023,37(1):231-251.
31 Zhan W,Ruan B,Dong H,et al.Isopsoralen suppresses receptor activator of nuclear factor kappa-β ligand-induced osteoclastogenesis by inhibiting the NF-κB signaling[J].Peer J,2023,11:e14560.
32 Zhu Z,Wang Z,Ma C,et al.Isopsoralen promotes osteogenic differentiation of human jawbone marrow mesenchymal cells through Notch signaling pathway[J].Ann Anat,2023,250:152156.
33 Zheng H,Feng H,Zhang W,et al.Targeting autophagy by natural product Ursolic acid for prevention and treatment of osteoporosis[J].Toxicol Appl Pharmacol,2020,409:115271.
34 Tan H,Zhao C,Zhu Q,et al.Ursolic acid isolated from the leaves of loquat(Eriobotrya japonica) inhibited osteoclast differentiation through targeting exportin 5[J].JAgric Food Chem,2019,67(12):3333-3340.
35 Xie BP,Shi LY,Li JP,et al.Oleanolic acid inhibits RANKL-induced osteoclastogenesis via ER alpha/mi R-503/RANK signaling pathway in RAW264.7 cells[J].Biomed Pharmacother,2019,117:109045.
36柴爽，马江涛，杨岩冰等.雌激素受体在骨疏康治疗绝经后骨质疏松症中的作用与机制[J].中国组织工程研究，2024,28(16):2574-2578.Chai S,Ma JT,Yang YB，等.The role and mechanism of estrogen receptor in the treatment ofpostmenopausal osteoporosis by Gushukang[J].Chin J Tissue Eng Res,2024,28(16):2574-2578.
37 Xia H,Liu J,Yang W,et al.Integrated strategy of network pharmacological prediction and experimental validation elucidate possible mechanism of Bu-Yang Herbs in treating postmenopausal osteoporosis via ESR1[J].Front Pharmacol,2021,12:654714.
38 Bajpai AK,Gu Q,Jiao Y,et al.Systems genetics and bioinformatics analyses using ESR1-correlated genes identify potential candidates underlying female bone development[J].Genomics,2024,116(1):110769.
39 Mukherjee A,Rotwein P.Selective signaling by Akt1controls osteoblast differentiation and osteoblast-mediated osteoclast development[J].Mol Cell Biol,2012,32(2):490-500.
40 Gao S,Chen B,Zhu Z,et al.PI3K-Akt signaling regulates BMP2-induced osteogenic differentiation of mesenchymal stem cells(MSCs):A transcriptomic landscape analysis[J].Stem Cell Res,2023,66:103010.
41 Wang Y,Liu L,Qu Z,et al.Tanshinone ameliorates glucocorticoid-induced bone loss via activation of AKT1signaling pathway[J].Front Cell Dev Biol,2022,10:878433.
42 Kisfali P,Polgár N,Sáfrány E,et al.Triglyceride level affecting shared susceptibility genes in metabolic syndrome and coronary artery disease[J].Curr Med Chem,2010,17(30):3533-3541.
43 Lecka-Czernik B,Gubrij I,Moerman EJ,et al.Inhibition of Osf2/Cbfa1 expression and terminal osteoblast differentiation by PPARgamma2[J].J Cell Biochem,1999,74(3):357-371.
44 Shockley KR,Lazarenko OP,Czernik PJ,et al.PPAR-gamma2 nuclear receptor controls multiple regulatory pathways of osteoblast differentiation from marrow mesenchymal stem cells[J].J Cell Biochem,2009,106(2):232-246.
45 Wan Y,Chong LW,Evans RM.PPAR-gamma regulates osteoclastogenesis in mice[J].Nat Med,2007,13(12):1496-1503.
46 Chen LS,Zhang M,Chen P,et al.The m(6)A demethylase FTO promotes the osteogenesis of mesenchymal stem cells by downregulating PPARG[J].Acta Pharmacol Sin,2022,43(5):1311-1323.
47 Marini F,Giusti F,Palmini G,et al.Role of Wnt signaling and sclerostin in bone and as therapeutic targets in skeletal disorders[J].Osteoporos Int,2023,34(2):213-238.
48 Delgado-Calle J,Sato AY,Bellido T.Role and mechanism of action of sclerostin in bone[J].Bone,2017,96:29-37.
49 Baroi S,Czernik PJ,Chougule A,et al.PPARG in osteocytes controls sclerostin expression,bone mass,marrow adiposity and mediates TZD-induced bone loss[J].Bone,2021,147:115913.
50 Li S,Jiang M,Wang L,et al.Combined chemotherapy with cyclooxygenase-2(COX-2) inhibitors in treating human cancers:Recent advancement[J].Biomed Pharmacother,2020,129:110389.
51罗鸣然，范文豪，李鑫等.靶向调控前列腺素内过氧化物合成酶2基因抑制MC3T3-E1细胞的增殖和成骨分化[J].中国组织工程研究，2022,26(12):1888-1893.Luo MR,Fan WH,Li X,et al.Inhibition of the proliferation and osteogenic differentiation of MC3T3-E1cells by targeting prostaglandin-endoperoxide synthase2[J].Chin J Tissue Eng Res,2022,26(12):1888-1893.
52 Lu L,Tian L.Postmenopausal osteoporosis coexisting with sarcopenia:The role and mechanisms of estrogen[J].J Endocrinol,2023,259(1):e230116[pii].
53 Hu HY,Zhang ZZ,Jiang XY,et al.Hesperidin anti-osteoporosis by regulating estrogen signaling pathways[J].Molecules,2023,28(19):6987.
54甘国兴，莫新民，李劲平等.壮骨止痛方石油醚提取A部位对去势雌鼠骨质疏松症的疗效及机理研究[J].中国中医基础医学杂志，2010,16(12):1169-1170,1185.Gan XG,Mo XM,Li JP,et al.Study on the Therapeutic effect and Mechanism of part an extracted from Zhuanggu Zhitong Formula on osteoporosis in ovariectomized female rats[J].Journal of Basic Chinese Medicine,2010,16(12):1169-1170,1185.
55彭昕，曾锦宁，张叔琦等.壮骨止痛方对体外培养成骨细胞Wnt/β-catenin信号通路的影响[J].中国中西医结合杂志，2017,37(9):1105-1108.Peng X,Zeng JN,Zhang SQ,et al.Effect of ZhuangguZhitong prescription on Wnt/β-catenin signaling pathway of ovariectomized rats with osteoporosis[J].Chin J Int Tradit Chin Western Med,2017,37(9):1105-1108.
56 Xiao Y,Zhao XP.Screening pathways and hub genes involved in osteoclastogenesis by gene expression analysis of circulating monocytes based on Gibbs sampling[J].Exp Ther Med,2019,17(4):2529-2534.
57 Uehara S,Udagawa N,Kobayashi Y.Regulation of osteoclast function via Rho-Pkn3-c-Src pathways[J].JOral Biosci,2019,61(3):135-140.
58 Xie G,Liu W,Lian Z,et al.Spleen tyrosine kinase(SYK) inhibitor PRT062607 protects against ovariectomy-induced bone loss and breast cancer-induced bone destruction[J].Biochem Pharmacol,2021,188:114579.
59 Matsubara T,Yasuda K,Mizuta K,et al.Tyrosine kinase src is a regulatory factor of bone homeostasis[J].Int J Mol Sci,2022,23(10):5508.
60张琦，李丹，杨芳等.“肾-精-髓-骨”系统的内涵与外延[J].中国中医基础医学杂志，2023,29(7):1055-1058.Zhang Q,Li D,Yang F,et al.Connotation and Extension of"Kidney-Essence-Marrow-Bone"System[J].J Basic Chin Med,2023,29(7):1055-1058.
基本信息:
DOI：10.13210/j.cnki.jhmu.20240425.001
中图分类号:R285
引用信息:
[1]陈诗淇,王意坚,陈瑶,等.基于网络药理学和分子对接探究壮骨止痛方治疗绝经后骨质疏松症的作用机制[J].海南医学院学报,2024,30(16):1246-1258.DOI:10.13210/j.cnki.jhmu.20240425.001.
基金信息:
国家自然科学基金资助项目（81973920）; 湖南省自然科学基金面上项目（2021JJ30494）; 湖南省卫生健康委项目（D202303107310）; 湖南省教育厅科学研究项目（20K092）; 湖南中医药大学校级研究生创新课题立项项目（2023CX170）~~
2024-04-25
2024-04-25
2024-04-25