[1]梁博程,史晓林,许超,等.基于中药系统药理学方法研究六味地黄丸治疗骨质疏松症的药效成分、作用靶点及作用特点[J].中医正骨,2019,31(04):1-7.
　LIANG Bocheng,SHI Xiaolin,XU Chao,et al.A study of pharmacodynamic components,action targets and characteristics of Liuwei Dihuang Wan(六味地黄丸)in treatment of osteoporosis using traditional chinese medicine systems pharmacology approach[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2019,31(04):1-7.
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基于中药系统药理学方法研究六味地黄丸治疗骨质疏松症的药效成分、作用靶点及作用特点()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第31卷
期数:
2019年04期
页码:
1-7
栏目:
基础研究
出版日期:
2019-04-30

文章信息/Info

Title:
A study of pharmacodynamic components,action targets and characteristics of Liuwei Dihuang Wan(六味地黄丸)in treatment of osteoporosis using traditional chinese medicine systems pharmacology approach
作者:
梁博程; 史晓林; 许超; 吴连国; 何滨; 李琰华; 李敏
(浙江中医药大学附属第二医院,浙江 杭州 310005)
Author(s):
LIANG Bocheng; SHI Xiaolin; XU Chao; WU Lianguo; HE Bin; LI Yanhua; LI Min
The Second Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310005,Zhejiang,China
关键词:
骨质疏松;  六味地黄丸;  药理作用;  基因本体论;  信号通路
Keywords:
osteoporosis;  Liuwei Dihuang Wan;  pharmacologic actions;  gene ontology;  signaling pathway
摘要:
目的:基于中药系统药理学方法探讨六味地黄丸治疗骨质疏松症(osteoporosis,OP)的药效成分、作用靶点及作用特点。方法:在中药综合数据库中检索六味地黄丸中的已知化学成分,然后使用中医药分子机制生物信息学分析工具预测六味地黄丸已知化学成分的作用靶点。通过检索DisGeNET数据库和人工阅读文献,查找OP相关靶基因和蛋白,通过与预测得到的作用靶点进行交叠,筛选六味地黄丸治疗OP的作用靶点。利用Cytoscape 3.7软件自带的BiNGO和ClueGO插件,分别对筛选出的靶点进行基因注释和信号通路富集分析。最后选取前期构建的绝经后OP疾病模型中的87个差异表达蛋白(基于血清蛋白质组学筛选的绝经后OP女性与绝经后骨量正常女性的血清差异表达蛋白),与筛选出的六味地黄丸治疗OP的作用靶点进行交叠,筛选六味地黄丸治疗OP的关键靶点,通过分析原始数据,确定相应的中药及其药效成分。结果:通过检索发现六味地黄丸化学成分130个,其中熟地黄化学成分8个、山药化学成分20个、山茱萸化学成分46个、泽泻化学成分21个、牡丹皮化学成分18个、茯苓化学成分21个; 茯苓与山茱萸、山茱萸与牡丹皮各含有1个相同化学成分,泽泻与山茱萸含有2个相同化学成分。根据六味地黄丸化学成分共预测到1169个作用靶点,其中君药熟地黄作用靶点42个,臣药山茱萸和山药作用靶点997个,佐药泽泻、牡丹皮和茯苓作用靶点共622个; 君药与臣药共有12个共同作用靶点,君药与佐药有14个共同作用靶点,臣药与佐药有472个共同作用靶点,君药、臣药、佐药共有6个共同作用靶点。经检索DisGeNET数据库和人工阅读文献,共检索到OP相关靶点687个,通过与预测到的六味地黄丸作用靶点交叠,共筛选出六味地黄丸治疗OP的作用靶点142个。靶点基因注释发现,六味地黄丸治疗OP的142个作用靶点主要参与了20种生物学过程,分布于8种细胞成分中,具有10种分子功能,其作用涉及骨重建、骨化、血管重塑、软骨发育、肌肉发育、骨骼发育、维生素D代谢、维生素应答、雌激素刺激应答、破骨细胞分化、成骨细胞分化、骨矿化、Wnt信号通路和Samd信号通路等生物学行为的调控。信号通路富集分析发现,119个信号通路被显著富集,破骨细胞分化、NF-κB信号通路和卵巢类固醇生成等24个功能组被显著富集,其中Wnt信号通路和与绝经后OP密切相关的雌激素调控信号通路等在24个功能组中广泛分布。通过将87个差异表达蛋白与筛选出的六味地黄丸治疗OP的作用靶点交叠,筛选出血管紧张素原、载脂蛋白E、Dickkopf相关蛋白3、RAS相关蛋白7a和蛋白质二硫键异构酶共5个关键靶点,对应山茱萸、山药、茯苓的6个药效成分,包括马兜铃酮、α-考绕咖烯、豆甾醇、尿囊素、鞘氨醇A及齿孔醇。结论:六味地黄丸治疗OP的药效成分包括山药、山茱萸和茯苓所含化合物马兜铃酮、α-考绕咖烯、豆甾醇、尿囊素、鞘氨醇A及齿孔醇,主要作用靶点包括血管紧张素原、载脂蛋白E、Dickkopf相关蛋白3、RAS相关蛋白7a和蛋白质二硫键异构酶。六味地黄丸治疗OP具有多靶点、多系统协同作用的特点。
Abstract:
Objective:To explore the pharmacodynamic components,action targets and characteristics of Liuwei Dihuang Wan(六味地黄丸,LWDHW)in treatment of osteoporosis(OP)using traditional Chinese medicine(TCM)systems pharmacology approach.Methods:The known chemical components of LWDHW were searched out from traditional Chinese medicine integrative database(TCMID),and the action targets of known chemical components of LWDHW were predicted by using a bioinformatics analysis tool for molecular mechanism of raditional Chinese medicine(BATMAN-TCM).The OP-related target genes and proteins were searched out through retrieving DisGeNET database and reading the articles,and the action targets of LWDHW for treatment of OP were selected out through overlapping the OP-related target genes and proteins with the predicted action targets.Gene annotation and signal pathway enrichment analysis were performed on the selected targets respectively by using BiNGO and ClueGO plug-ins in Cytoscape 3.7 software.Eighty-seven proteins,which differentially expressed in serum of females with PMOP and postmenopausal females with normal bone mass,were selected out from the PMOP models to overlap with the selected action targets of LWDHW for treatment of OP,and the key targets of LWDHW for treatment of OP were selected out finally.The corresponding TCMs and their pharmacodynamic components were determined through analyzing the original data.Results:One hundred and thirty chemical components of LWDHW were found out through retrieving TCMID,including the chemical components of radix rehmanniae praeparata(8),rhizoma dioscoreae(20),fructus corni(46),rhizoma alismatis(21),cortex moutan radicis(18)and poria cocos(21).One identical chemical component was found in poria cocos and fructus corni and in fructus corni and cortex moutan radicis respectively,and two identical chemical components were found in rhizoma alismatis and fructus corni.One thousand one hundred and sixty-nine action targets were predicted according to the chemical components of LWDHW,including the action targets of radix rehmanniae praeparata(sovereign drug,42),fructus corni and rhizoma dioscoreae(minister drug,997)and rhizoma alismatis,cortex moutan radicis and poria cocos(assistant drug,622).The common action targets were found in sovereign drug and minister drug(12),sovereign drug and assistant drug(14),minister drug and assistant drug(472)and sovereign drug,minister drug and assistant drug(6).Six hundred and eighty-seven OP-related targets were searched out through retrieving DisGeNET database and reading the articles,and 142 action targets of LWDHW for treatment of OP were selected out by overlapping the OP-related targets with the predicted action targets of LWDHW.The results of gene annotation for the targets demonstrated that the 142 action targets of LWDHW for treatment of OP were mainly involved in 20 kinds of biological processes,distributed in 8 kinds of cell components and had 10 kinds of molecular functions,and played a role in the regulation of biological behaviors such as bone reconstruction,ossification,vascular remodeling,cartilage development,muscle development,bone development,vitamin D metabolism,response to vitamin,response to estrogen stimulation,osteoclast differentiation,osteoblast differentiation,bone mineralization,Wnt signaling pathway and Samd signaling pathway.The results of signal pathway enrichment analysis of the targets demonstrated that 119 signaling pathways were significantly enriched,and 24 functional groups including osteoclast differentiation,NF-κB signaling pathway and ovarian steroidogenesis were significantly enriched,and Wnt signaling pathway and PMOP-related estrogen regulation signaling pathway were widely distributed in the 24 functional groups.Five key targets,including angiotensinogen(AGT),apolipoprotein E(APOE),Dickkopf-related protein 3(DKK3),RAB7A and protein disulfide isomerase(PDI),were selected out through overlapping the 87 differentially expressed proteins with the selected action targets of LWDHW for treatment of OP,and they corresponded to the 6 pharmacodynamic components of fructus corni,rhizoma dioscoreae and poria cocos,including aristolone,alpha-corocalene,stigmasterol,allantoin,sphingosine A and eburicol.Conclusion:The pharmacodynamic components of LWDHW for treatment of OP include aristolone,alpha-corocalene,stigmasterol,allantoin,sphingosine A and eburicol,which are contained in rhizoma dioscoreae,fructus corni and poria cocos,and their main action targets include AGT,APOE,DKK3,RAB7A and PDI.LWDHW is characterized by synergistic effect of multiple targets and multiple systems in treatment of OP.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金项目(81803902) 通讯作者:李敏 E-mail:liminist@163.com(收稿日期:2018-12-28 本文编辑:李晓乐)
更新日期/Last Update: 2019-10-08