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Article(id=1218551240372441120, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1218551237453201879, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2018-0506, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1527523200000, receivedDateStr=2018-05-29, revisedDate=1530633600000, revisedDateStr=2018-07-04, acceptedDate=null, acceptedDateStr=null, onlineDate=1768454855292, onlineDateStr=2026-01-15, pubDate=1539273600000, pubDateStr=2018-10-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768454855292, onlineIssueDateStr=2026-01-15, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768454855292, creator=13701087609, updateTime=1768454855292, updator=13701087609, issue=Issue{id=1218551237453201879, tenantId=1146029695717560320, journalId=1189982191388893191, year='2018', volume='53', issue='10', pageStart='1583', pageEnd='1760', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768454854596, creator=13701087609, updateTime=1768457142735, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1218560834666680483, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1218551237453201879, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1218560834670874788, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1218551237453201879, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1670, endPage=1679, ext={EN=ArticleExt(id=1218551240980615266, articleId=1218551240372441120, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=Mechanism of detoxification of Chebulae Fructus against Aconiti kusnezoffii radix toxicity based on network pharmacology, columnId=1218551213583421639, journalTitle=Acta Pharmaceutica Sinica, columnName=Pharmacology, runingTitle=null, highlight=null, articleAbstract=

The mechanism of detoxification of Chebulae Fructus against Aconiti kusnezoffii radix toxicity, which was known as Mongolian medical theory, was studied by establishing network of active components-targets-pathways of detoxification and enrichment analysis of targets and pathways based on network pharmacology. Firstly, the targets of active components collected from TCMSP and TCM Database@Taiwan were obtained through SwissTargetPrediction compared with disease targets from OMIM, TTD, DiGSeE. Then, the target enrichment analysis of GO functional annotations and KEGG pathways and protein function were analyzed by Metascape, furthermore, the action between main active ingredients and targets was assessed by SystemsDock Web Site. At last, the Cytoscape was used to construct the network of active components-targets-pathways. In conclusion, there were 15 components and 40 targets related to the cardiotoxicity caused by Aconiti kusnezoffii radix. Furthermore, Chebulae Fructus could regulate cardiac function to detoxify the toxicity by Aconiti kusnezoffii radix through the biological process of negative regulation of blood vessel diameter, regulation of ion transport circulatory system process, muscle contraction inorganic ion homeostasis and the pathways of neuroactive ligand-receptor interaction, calcium signaling pathway, adrenergic signaling in cardiomyocytes, etc.

, correspAuthors=Ya TU, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2018 Acta Pharmaceutica Sinica. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Jian-liang LI, Hui LIANG, Shu-zhen CAI, Zhi-yong LI, Ya TU), CN=ArticleExt(id=1218551247083328303, articleId=1218551240372441120, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=基于网络药理学探讨蒙药诃子解草乌心脏毒的机制研究, columnId=1218551241983054033, journalTitle=药学学报, columnName=药理学, runingTitle=null, highlight=null, articleAbstract=

利用网络药理学的方法，建立诃子解草乌毒性的活性成分-作用靶点-通路的网络图，对作用靶点的基因功能和通路进行富集分析，探讨传统蒙医理论诃子解草乌心脏毒性的作用机制。通过TCMSP数据库、台湾中医药资料库获得诃子主要活性成分，采用SwissTargetPrediction预测活性成分的潜在靶点并于OMIM、TTD、DiGSeE三个疾病靶点数据进行比对，获得诃子解草乌心脏毒性的作用靶点。采用Metascape数据库平台对作用靶点的GO（Gene ontology）注释和KEGG通路进行富集分析和蛋白功能归属，并利用SystemsDock分子对接网站对诃子成分与靶点进行分子对接验证。最后通过Cytoscape构建诃子解草乌毒“活性成分-作用靶点-通路”网络图，最终发现诃子15个活性成分、40个作用靶点与其缓解草乌引起的心脏毒性相关。结果表明，诃子主要涉及配体受体结合、肌肉收缩、循环系统进程和细胞内外离子交换等生物过程并通过调节神经活性配体受体相互作用、钙离子信号通路、心肌细胞肾上腺素能信号等通路调节心脏功能进一步缓解草乌引起的心脏毒性。

, correspAuthors=图雅, authorNote=null, correspAuthorsNote=
* 图雅, Tel:86-10-64089691, E-mail:
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Development Research Center of TCM, China Academy of Traditional Chinese Medicine, Beijing 100700, China), AuthorCompanyExt(id=1218970770798203594, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, companyId=1218970770760454850, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=5.中国中医科学院中医药研究发展中心, 北京 100700)])], figs=[ArticleFig(id=1218970777530060941, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=doKwUz3zWjXI5O4J6X6R3g==, figureFileBig=G5WB8QWz9cQV9aH+TLpAJQ==, tableContent=null), ArticleFig(id=1218970777685250198, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Figure 1, caption=

Enrichment analysis of potential targets from main active ingredients of Chebulae Fructus against Aconiti kusnezoffii radix toxicity. Analysis of Gene Ontology terms for biological process (A), molecular function (B) and cellular component (C); Analysis of KEGG pathways (D)

, figureFileSmall=doKwUz3zWjXI5O4J6X6R3g==, figureFileBig=G5WB8QWz9cQV9aH+TLpAJQ==, tableContent=null), ArticleFig(id=1218970777861410984, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=7+8oJPUYbhAbMsfXoGO/rg==, figureFileBig=kOZApBDlPcR4Ibdrv4w+Mg==, tableContent=null), ArticleFig(id=1218970777987240119, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Figure 2, caption=

Components-targets-pathways network of Chebulae Fructus against Aconiti kusnezoffii radix toxicity

, figureFileSmall=7+8oJPUYbhAbMsfXoGO/rg==, figureFileBig=kOZApBDlPcR4Ibdrv4w+Mg==, tableContent=null), ArticleFig(id=1218970778079514817, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=7aBBwt7PcLZDWShXXlESlw==, figureFileBig=/Vb79UZ7uD4GIn/W0x5iUw==, tableContent=null), ArticleFig(id=1218970778196955347, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Figure 3, caption=

The docking scores (pKd/pKi) of 4 target from 15 active ingredients in Chebulae Fructus compared to lidocaine

, figureFileSmall=7aBBwt7PcLZDWShXXlESlw==, figureFileBig=/Vb79UZ7uD4GIn/W0x5iUw==, tableContent=null), ArticleFig(id=1218970778322784481, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Name Chemical formula 2D structure
H1 Delta(7)-stigmastenol C29H50O
H2 Glucosol C30H48O4
H3 Maslinic acid C30H48O4
H4 Arjungenin C30H48O6
H5 Arjunolic acid C30H48O5
H6 Ellagic acid C14H6O8
H7 Chebulic acid C14H12O11
H8 Corilagin C27H22O18
H9 Tannic acid C27H24O18
H10 Pentagalloyl glucose C41H32O26
H11 Sennoside A C42H38O20
H12 Sennoside E C44H38O23
H13 Cheilanthifoline C19H19NO4
H14 Peraksine C19H22N2O2
H15 Quinidine sulfate C20H24N2O2
), ArticleFig(id=1218970778452807917, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Table 1, caption=

Main active ingredients from Chebulae Fructus

, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Name Chemical formula 2D structure
H1 Delta(7)-stigmastenol C29H50O
H2 Glucosol C30H48O4
H3 Maslinic acid C30H48O4
H4 Arjungenin C30H48O6
H5 Arjunolic acid C30H48O5
H6 Ellagic acid C14H6O8
H7 Chebulic acid C14H12O11
H8 Corilagin C27H22O18
H9 Tannic acid C27H24O18
H10 Pentagalloyl glucose C41H32O26
H11 Sennoside A C42H38O20
H12 Sennoside E C44H38O23
H13 Cheilanthifoline C19H19NO4
H14 Peraksine C19H22N2O2
H15 Quinidine sulfate C20H24N2O2
), ArticleFig(id=1218970778603802878, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Gene symbol Description
1 OPRM1 Opioid receptor mu 1
2 SLC6A2 Solute carrier family 6 member 2
3 TNNC1 Troponin C1, slow skeletal and cardiac type
4 HMGCR 3-Hydroxy-3-methylglutaryl-coa reductase
5 DRD2 Dopamine receptor D2
6 SLC6A4 Solute carrier family 6 member 4
7 KCNA1 Potassium voltage-gated channel subfamily A member 1
8 KCNA4 Potassium voltage-gated channel subfamily A member 4
9 KCNA3 Potassium voltage-gated channel subfamily A member 3
10 KCNA5 Potassium voltage-gated channel subfamily A member 5
11 MMP2 Matrix metallopeptidase 2
12 GSTM1 Glutathione S-transferase mu 1
13 PGR Progesterone receptor
14 HTR1A 5-Hydroxytryptamine receptor 1A
15 BCHE Butyrylcholinesterase
16 HTR3A 5-Hydroxytryptamine receptor 3A
17 LACTB Lactamase beta
18 SCN5A Sodium voltage-gated channel alpha subunit 5
19 AR Androgen receptor
20 F10 Coagulation factor X
21 ESR1 Estrogen receptor 1
22 ABCB1 ATP binding cassette subfamily B member 1
23 ESR2 Estrogen receptor 2
24 TNNI3 Troponin I3, cardiac type
25 ABCB4 ATP binding cassette subfamily B member 4
26 TNNT2 Troponin T2, cardiac type
27 TNNT1 Troponin T1, slow skeletal type
28 CHRM4 Cholinergic receptor muscarinic 4
29 CHRM2 Cholinergic receptor muscarinic 2
30 CHRM1 Cholinergic receptor muscarinic 1
31 AKR1B10 Aldo-keto reductase family 1 member B10
32 F2 Coagulation factor II, thrombin
33 ADRA1B Adrenoceptor alpha 1B
34 HSD11B1 Hydroxysteroid 11-beta dehydrogenase 1
35 ADRA1A Adrenoceptor alpha 1A
36 CA2 Carbonic anhydrase 2
37 KCNH2 Potassium voltage-gated channel subfamily H member 2
38 ADRA1D Adrenoceptor alpha 1D
39 OPRD1 Opioid receptor delta 1
40 HTR2A 5-Hydroxytryptamine receptor 2A
), ArticleFig(id=1218970778763186448, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Table 2, caption=

Protein function potential targets from main active ingredients of Chebulae Fructus against Aconiti kusnezoffii radix toxicity

, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Gene symbol Description
1 OPRM1 Opioid receptor mu 1
2 SLC6A2 Solute carrier family 6 member 2
3 TNNC1 Troponin C1, slow skeletal and cardiac type
4 HMGCR 3-Hydroxy-3-methylglutaryl-coa reductase
5 DRD2 Dopamine receptor D2
6 SLC6A4 Solute carrier family 6 member 4
7 KCNA1 Potassium voltage-gated channel subfamily A member 1
8 KCNA4 Potassium voltage-gated channel subfamily A member 4
9 KCNA3 Potassium voltage-gated channel subfamily A member 3
10 KCNA5 Potassium voltage-gated channel subfamily A member 5
11 MMP2 Matrix metallopeptidase 2
12 GSTM1 Glutathione S-transferase mu 1
13 PGR Progesterone receptor
14 HTR1A 5-Hydroxytryptamine receptor 1A
15 BCHE Butyrylcholinesterase
16 HTR3A 5-Hydroxytryptamine receptor 3A
17 LACTB Lactamase beta
18 SCN5A Sodium voltage-gated channel alpha subunit 5
19 AR Androgen receptor
20 F10 Coagulation factor X
21 ESR1 Estrogen receptor 1
22 ABCB1 ATP binding cassette subfamily B member 1
23 ESR2 Estrogen receptor 2
24 TNNI3 Troponin I3, cardiac type
25 ABCB4 ATP binding cassette subfamily B member 4
26 TNNT2 Troponin T2, cardiac type
27 TNNT1 Troponin T1, slow skeletal type
28 CHRM4 Cholinergic receptor muscarinic 4
29 CHRM2 Cholinergic receptor muscarinic 2
30 CHRM1 Cholinergic receptor muscarinic 1
31 AKR1B10 Aldo-keto reductase family 1 member B10
32 F2 Coagulation factor II, thrombin
33 ADRA1B Adrenoceptor alpha 1B
34 HSD11B1 Hydroxysteroid 11-beta dehydrogenase 1
35 ADRA1A Adrenoceptor alpha 1A
36 CA2 Carbonic anhydrase 2
37 KCNH2 Potassium voltage-gated channel subfamily H member 2
38 ADRA1D Adrenoceptor alpha 1D
39 OPRD1 Opioid receptor delta 1
40 HTR2A 5-Hydroxytryptamine receptor 2A
), ArticleFig(id=1218970778918375714, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Gene symbol Protein function
1 OPRM1 G-protein coupled receptors/GPCRs excl olfactory receptors; transporters
2 SLC6A2 Transporters/electrochemical potential-driven transporters
3 TNNC1 Null
4 HMGCR Enzymes/enzyme proteins/oxidoreductases; transporters/electrochemical potential-driven transporters
5 DRD2 G-Protein coupled receptors/GPCRs excl olfactory receptors; transporters
6 SLC6A4 Transporters/electrochemical potential-driven transporters
7 KCNA1 Voltage-gated ion channels/voltage-gated potassium channels
8 KCNA4 Voltage-gated ion channels/voltage-gated potassium channels
9 KCNA3 Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated potassium channels
10 KCNA5 Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated potassium channels
11 MMP2 Enzymes/[enzyme proteins/hydrolases, peptidases/metallopeptidases]
12 GSTM1 Enzymes/enzyme proteins/transferases
13 PGR Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
14 HTR1A G-Protein coupled receptors/[GPCRs excl olfactory receptors, serotonin receptors]
15 BCHE Enzymes/enzyme proteins/hydrolases
16 HTR3A Transporters/transporter channels and pores
17 LACTB Enzymes/peptidases/serine-type peptidases
18 SCN5A Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated sodium channels
19 AR Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
20 F10 Enzymes/[enzyme proteins/hydrolases, peptidases/serine-type peptidases]
21 ESR1 Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
22 ABCB1 CD markers; enzymes/enzyme proteins/hydrolases; transporters/primary active transporters
23 ESR2 Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
24 TNNI3 Null
25 ABCB4 Enzymes/enzyme proteins/hydrolases; transporters/primary active transporters
26 TNNT2 Null
27 TNNT1 Null
28 CHRM4 G-Protein coupled receptors/GPCRs excl olfactory receptors
29 CHRM2 G-Protein coupled receptors/GPCRs excl olfactory receptors
30 CHRM1 G-Protein coupled receptors/GPCRs excl olfactory receptors
31 AKR1B10 Null
32 F2 Enzymes/[enzyme proteins/hydrolases, peptidases/serine-type peptidases]
33 ADRA1B G-Protein coupled receptors/GPCRs excl olfactory receptors
34 HSD11B1 Enzymes/enzyme proteins/oxidoreductases
35 ADRA1A G-Protein coupled receptors/GPCRs excl olfactory receptors
36 CA2 Enzymes/enzyme proteins/lyases
37 KCNH2 Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated potassium channels
38 ADRA1D G-Protein coupled receptors/GPCRs excl olfactory receptors
39 OPRD1 G-Protein coupled receptors/GPCRs excl olfactory receptors
40 HTR2A G-Protein coupled receptors/[GPCRs excl olfactory receptors, serotonin receptors]
), ArticleFig(id=1218970779052593451, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Table 3, caption=

Protein function potential targets from main active ingredients of Chebulae Fructus against Aconiti kusnezoffii radix toxicity

, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Gene symbol Protein function
1 OPRM1 G-protein coupled receptors/GPCRs excl olfactory receptors; transporters
2 SLC6A2 Transporters/electrochemical potential-driven transporters
3 TNNC1 Null
4 HMGCR Enzymes/enzyme proteins/oxidoreductases; transporters/electrochemical potential-driven transporters
5 DRD2 G-Protein coupled receptors/GPCRs excl olfactory receptors; transporters
6 SLC6A4 Transporters/electrochemical potential-driven transporters
7 KCNA1 Voltage-gated ion channels/voltage-gated potassium channels
8 KCNA4 Voltage-gated ion channels/voltage-gated potassium channels
9 KCNA3 Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated potassium channels
10 KCNA5 Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated potassium channels
11 MMP2 Enzymes/[enzyme proteins/hydrolases, peptidases/metallopeptidases]
12 GSTM1 Enzymes/enzyme proteins/transferases
13 PGR Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
14 HTR1A G-Protein coupled receptors/[GPCRs excl olfactory receptors, serotonin receptors]
15 BCHE Enzymes/enzyme proteins/hydrolases
16 HTR3A Transporters/transporter channels and pores
17 LACTB Enzymes/peptidases/serine-type peptidases
18 SCN5A Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated sodium channels
19 AR Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
20 F10 Enzymes/[enzyme proteins/hydrolases, peptidases/serine-type peptidases]
21 ESR1 Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
22 ABCB1 CD markers; enzymes/enzyme proteins/hydrolases; transporters/primary active transporters
23 ESR2 Nuclear receptors; transcription factors/zinc-coordinating DNA-binding domains
24 TNNI3 Null
25 ABCB4 Enzymes/enzyme proteins/hydrolases; transporters/primary active transporters
26 TNNT2 Null
27 TNNT1 Null
28 CHRM4 G-Protein coupled receptors/GPCRs excl olfactory receptors
29 CHRM2 G-Protein coupled receptors/GPCRs excl olfactory receptors
30 CHRM1 G-Protein coupled receptors/GPCRs excl olfactory receptors
31 AKR1B10 Null
32 F2 Enzymes/[enzyme proteins/hydrolases, peptidases/serine-type peptidases]
33 ADRA1B G-Protein coupled receptors/GPCRs excl olfactory receptors
34 HSD11B1 Enzymes/enzyme proteins/oxidoreductases
35 ADRA1A G-Protein coupled receptors/GPCRs excl olfactory receptors
36 CA2 Enzymes/enzyme proteins/lyases
37 KCNH2 Transporters/transporter channels and pores; voltage-gated ion channels/voltage-gated potassium channels
38 ADRA1D G-Protein coupled receptors/GPCRs excl olfactory receptors
39 OPRD1 G-Protein coupled receptors/GPCRs excl olfactory receptors
40 HTR2A G-Protein coupled receptors/[GPCRs excl olfactory receptors, serotonin receptors]
), ArticleFig(id=1218970779182616886, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
NO. Gene symbol Relevance score
1 SCN5A 19.57
2 CYP3A4 12.72
3 F2 11.04
4 CYP1A2 10.52
5 HBG2 10.36
6 IL5 8.5
7 SLC17A5 8.1
8 SCN9A 8.03
9 EGFR 7.79
10 ITGAM 7.73
), ArticleFig(id=1218970779312640328, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1218551240372441120, language=CN, label=Table 4, caption=

Top 10 genes related to lidocaine from GeneCards

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NO. Gene symbol Relevance score
1 SCN5A 19.57
2 CYP3A4 12.72
3 F2 11.04
4 CYP1A2 10.52
5 HBG2 10.36
6 IL5 8.5
7 SLC17A5 8.1
8 SCN9A 8.03
9 EGFR 7.79
10 ITGAM 7.73
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基于网络药理学探讨蒙药诃子解草乌心脏毒的机制研究
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李建良 1 , 梁慧 2 , 蔡淑珍 3 , 李志勇 4 , 图雅 5, *
药学学报 | 药理学 2018,53(10): 1670-1679
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药学学报 | 药理学 2018, 53(10): 1670-1679
基于网络药理学探讨蒙药诃子解草乌心脏毒的机制研究
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李建良1, 梁慧2, 蔡淑珍3, 李志勇4, 图雅5, *
作者信息
  • 1.中国中医科学院医学实验中心, 北京 100700
  • 2.内蒙古医科大学蒙医药学院, 内蒙古 呼和浩特 010110
  • 3.内蒙古民族大学蒙医药学院, 内蒙古 通辽 028000
  • 4.中央民族大学药学院, 北京 100081
  • 5.中国中医科学院中医药研究发展中心, 北京 100700

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* 图雅, Tel:86-10-64089691, E-mail:
Mechanism of detoxification of Chebulae Fructus against Aconiti kusnezoffii radix toxicity based on network pharmacology
Jian-liang LI1, Hui LIANG2, Shu-zhen CAI3, Zhi-yong LI4, Ya TU5, *
Affiliations
  • 1. Medical Research Center, China Academy of Traditional Chinese Medicine, Beijing 100700, China
  • 2. Mongolian Medicine College, Inner Mongolia Medical University, Huhehaote 010110, China
  • 3. Mongolian Medicine College, Inner Mongolia University of Nationalities, Tongliao 028000, China
  • 4. Pharmacy College of Minzu University of China, Beijing 100081, China
  • 5. Development Research Center of TCM, China Academy of Traditional Chinese Medicine, Beijing 100700, China
出版时间: 2018-10-12 doi: 10.16438/j.0513-4870.2018-0506
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摘要
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利用网络药理学的方法，建立诃子解草乌毒性的活性成分-作用靶点-通路的网络图，对作用靶点的基因功能和通路进行富集分析，探讨传统蒙医理论诃子解草乌心脏毒性的作用机制。通过TCMSP数据库、台湾中医药资料库获得诃子主要活性成分，采用SwissTargetPrediction预测活性成分的潜在靶点并于OMIM、TTD、DiGSeE三个疾病靶点数据进行比对，获得诃子解草乌心脏毒性的作用靶点。采用Metascape数据库平台对作用靶点的GO（Gene ontology）注释和KEGG通路进行富集分析和蛋白功能归属，并利用SystemsDock分子对接网站对诃子成分与靶点进行分子对接验证。最后通过Cytoscape构建诃子解草乌毒“活性成分-作用靶点-通路”网络图，最终发现诃子15个活性成分、40个作用靶点与其缓解草乌引起的心脏毒性相关。结果表明，诃子主要涉及配体受体结合、肌肉收缩、循环系统进程和细胞内外离子交换等生物过程并通过调节神经活性配体受体相互作用、钙离子信号通路、心肌细胞肾上腺素能信号等通路调节心脏功能进一步缓解草乌引起的心脏毒性。

诃子  /  草乌毒性  /  蒙医理论  /  网络药理学  /  作用机制

The mechanism of detoxification of Chebulae Fructus against Aconiti kusnezoffii radix toxicity, which was known as Mongolian medical theory, was studied by establishing network of active components-targets-pathways of detoxification and enrichment analysis of targets and pathways based on network pharmacology. Firstly, the targets of active components collected from TCMSP and TCM Database@Taiwan were obtained through SwissTargetPrediction compared with disease targets from OMIM, TTD, DiGSeE. Then, the target enrichment analysis of GO functional annotations and KEGG pathways and protein function were analyzed by Metascape, furthermore, the action between main active ingredients and targets was assessed by SystemsDock Web Site. At last, the Cytoscape was used to construct the network of active components-targets-pathways. In conclusion, there were 15 components and 40 targets related to the cardiotoxicity caused by Aconiti kusnezoffii radix. Furthermore, Chebulae Fructus could regulate cardiac function to detoxify the toxicity by Aconiti kusnezoffii radix through the biological process of negative regulation of blood vessel diameter, regulation of ion transport circulatory system process, muscle contraction inorganic ion homeostasis and the pathways of neuroactive ligand-receptor interaction, calcium signaling pathway, adrenergic signaling in cardiomyocytes, etc.

Chebulae Fructus  /  Aconiti kusnezoffii radix toxicity  /  Mongolian medical theory  /  network pharmacology  /  mechanism
李建良, 梁慧, 蔡淑珍, 李志勇, 图雅. 基于网络药理学探讨蒙药诃子解草乌心脏毒的机制研究. 药学学报, 2018 , 53 (10) : 1670 -1679 . DOI: 10.16438/j.0513-4870.2018-0506
Jian-liang LI, Hui LIANG, Shu-zhen CAI, Zhi-yong LI, Ya TU. Mechanism of detoxification of Chebulae Fructus against Aconiti kusnezoffii radix toxicity based on network pharmacology[J]. Acta Pharmaceutica Sinica, 2018 , 53 (10) : 1670 -1679 . DOI: 10.16438/j.0513-4870.2018-0506
正文
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蒙医药是蒙古族在长期的生产实践中同疾病抗争的经验积累和总结, 是我国四大民族医药之一; 其以三根学说、七素学说、寒热理论、脏腑理论、六基病症理论为基础, 形成了独特理论体系。临床上蒙医以成药应用为主, 药材多生用, 但是对于有毒或性质剧烈的药材, 为降低毒性, 改善药性则需进行必要的加工炮制。而蒙药的炮制经过了历代临床用药实践积累了丰富的经验, 同时也离不开蒙医药理论的指导[1, 2]。
蒙药草乌毛茛科植物北乌头(Aconitum kusnezoffii Reichb.)的干燥块根, 味辛、性温、轻。具有杀“粘”、止痛、燥“协日乌素”的功效, 临床用于治疗“痛风病”、“游痛病”、关节疼痛、偏瘫和心“赫依”等病症[3]。其有大毒[4], 需炮制后入药。草乌的炮制方法有使用酸奶、童便(尿)、黄酒、甘草汤、诃子汤炮制等[5], 其中, 诃子汤炮制为草乌主要方法, 也是特色方法, 现有的《内蒙古蒙成药标准》中已明确提出了用诃子制草乌。
当前对民族医药的研究重点应是以抓住民族医药形成过程的理论经验特质与药物采、制、用等特色, 将承载于民族药上的民族医药理论与经验用现代科学方法予以展现, 证明其有效性、科学性, 或阐释一种新的发现与补充; 因此, 当今的民族药研究亦应遵循“药效物质基本清楚、作用机制基本清楚和民族医药理论与经验特征基本清楚”的基本原则[6]。本文将采用网络药理学研究策略[7], 从诃子的物质基础出发, 着眼于诃子直接缓解草乌引起的心律不齐的心脏毒性, 从整体性的角度为诃子解草乌心脏毒性这一理论提供分子机制, 也为后续研究提供一定的理论基础。
材料与方法
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诃子化合物的获取和筛选  通过TCMSP数据库(http://lsp.nwu.edu.cn/tcmsp.php)、台湾中医药资料库(http://tcm.cmu.edu.tw/zh-tw/index.php)对诃子的化学成分进行搜集, 根据文献记载的诃子主要成分以及课题组前期研究基础整理出诃子能够改善草乌引起的心律不齐的活性成分。
诃子活性化合物的靶点预测  利用PubChem (https://pubchem.ncbi.nlm.nih.gov/)数据库, 将所有的化合物都转化为标准的Canonical SMILES格式, 将SMILES格式文件导入SwissTargetPrediction (http://www.swisstargetprediction.ch/)平台, 设置属性为“homo sapiens”, 预测活性化合物的潜在靶点并根据文献报道补充未预测到的活性化合物的已知靶点[8]。
诃子解草乌心脏毒性靶点的搜集  以“arrhythmia”为关键词, 通过OMIM (http://www.omim.org/)、TTD (http://bidd.nus.edu.sg/BIDDDatabases/TTD/TTD.asp及DiGSeE (http://210.107.182.61/geneSearch/)三个疾病靶点网站搜集的心律不齐的基因, 去除重复靶点基因, 与上一步获得的靶点进行比对, 最终获得诃子活性成分解草乌心脏毒性的潜在作用靶点。
生物过程与通路的富集分析  Metascape平台是一个基因注释分析数据库, 对输入基因进行生物过程和通路的富集分析。同时, Metascape平台不仅能够提供最新的数据库资源和全面的功能注释, 而且能够做出新的预测和通过已有的数据库对实验进行验证[9]。将诃子解心脏草乌毒性靶点放入Metascape平台, 提交后将输入物种和分析物种均选择为“H. sapiens”, 设置P < 0.01, 对诃子作用靶点进行GO注释分析及KEGG通路分析, 保存结果, 并按照每个条目涉及的靶点数目进行排序, 筛选排名靠前的生物过程和通路, 采用GraphPad Prism 7.0软件对结果可视化。
作用靶点蛋白功能归属   Metascape平台对于基因的注释分析不仅能对输入基因进行生物过程和通路的富集分析, 还能够收集其功能以及靶点类型。采用上述方法, 将诃子预测靶点导入Metascape数据库中, 获得其靶点功能信息(protein function)。
诃子解草乌毒“活性成分-靶点-通路”网络构建与分析   将诃子的活性成分、解草乌心脏毒作用的靶点和代谢通路的数据导入Cytoscape3.5.1软件, 采用Merge工具构建诃子解草乌毒“成分-靶点-通路”网络模型。
分子对接验证  SystemsDock分子对接网站(http://systemsdock.unit.oist.jp/iddp/home/index), 主要基于网络药理学的预测和分析, 并在复杂分子网络上通过对接模拟和分子通路图对配体选择性和配体动作的解释综合特征以评估蛋白质-配体结合潜能[10]。以“lidocaine”为关键词, 于GeneCards (https://www.genecards.org/)平台进行检索, 以“score”为参数进行排序, 获得相关性靠前的靶点并与诃子活性成分解草乌心脏毒性的潜在作用靶点进行比对, 将比对结果靶点和诃子活性成分上传至SystemsDock分子对接网站获得docking scores (pKd/pKi)进行分析, 以评价诃子活性成分与靶点之间的结合活性, 并采用GraphPad Prism 7.0软件对结果可视化。
结果
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1 诃子活性成分的筛选
诃子的化合物结构类型多, 通过TCMSP、台湾中医药资料库两个化合物数据库收集, 共收集54个化合物, 主要含有鞣质、三萜类、奎宁酸、诃子素、生物碱等。本课题组前期从化学成分的角度, 认为诃子中大量鞣质与草乌生物碱生成难溶性盐, 在体内减缓吸收而进一步降低毒性[11], 因此, 笔者通过TCMSP数据库筛选出DL (drug-like) ≥0.15的化合物并添加了含量较高的鞣质, 共有15个化合物(表 1), 分别为四环三萜(1个)、五环三萜(4个)、鞣质(7个)、生物碱(3个)。
2 靶点预测
基于SwissTargetPrediction平台来预测活性化合物的潜在靶点, 共获得175个基因靶点, 删除重复项, 共获得97个潜在靶点。通过和OMIM、TTD以及DiGSeE疾病数据库中对于心律不齐和心脏毒性相关基因的比对, 得到化合物潜在靶点和疾病靶点的交集, 最终筛选出40个可能与诃子解草乌心脏毒性作用相关的靶点, 见表 2。
3 基因功能和通路富集分析
使用Metascape数据库对40个诃子解草乌毒的潜在靶点进行GO注释分析和KEGG pathway分析, 设置阈值P < 0.01, 筛选出靠前的GO注释结果和KEGG通路, 并采用GraphPad Prism 7.0绘制热图, 结果见图 1。
GO注释分析指的是通过相同的语义描述基因生物学功能的项目, 由分子功能(molecular function, MF)、生物过程(biological process, BP)和细胞组成(cellular component, CC)三个部分组成。GO注释的数据量大, 其中, 生物过程(图 1A)中排名靠前的有系统进程调节、磷脂酶C-激活性G蛋白偶联受体信号通路、生物碱应答、铵转运、血管直径负调节; 分子功能分析(图 1B)主要是d延迟整流钾通道活性、G蛋白偶联胺受体活性、无机阳离子跨膜转运蛋白活性、阳离子跨膜转运蛋白活性、金属离子跨膜转运活性靠前, 主要涉及离子通道、膜内外离子转运活性; 而在细胞组成(图 1C)分析中靠前的主要是轴突、膜筏、膜微区、膜区、肌钙蛋白复合物、突触, 主要集中在神经元和细胞膜上。
通过Metascape数据库中的KEGG pathway富集分析, 诃子解草乌毒的靶点共涉及17条通路(图 1D), 主要是神经活性配体受体相互作用、钙离子信号通路、心肌细胞肾上腺素能信号通路、cAMP信号通路、雌激素信号通路等。
4 作用靶点蛋白功能归属
将40个作用靶点导入Metascape数据库, 获得靶点对应的蛋白功能, 结果见表 3。结果表明, 在诃子解草乌心脏毒性的过程中, 有转录因子、受体、酶和离子通道等物质的参与。
5 诃子解草乌毒“活性成分-靶点-通路”网络构建
将诃子的活性成分、作用靶点和KEGG通路信息导入Cytoscape软件中构建“活性成分-靶点-通路”网络。在输出的网络(图 2)中, 以不同颜色的节点表示活性成分、靶点以及作用通路, 红色圆形节点代表诃子活性成分, 紫色菱形节点代表作用靶点, 而绿色矩形代表作用通路。若某一靶点为活性化合物的潜在靶点, 则以边相连, 若某一蛋白对某条通路具有调节作用, 也通过边连接。从图中可以看出同一活性成分对应多个作用靶点和多个活性成分对应同一靶点的关系, 体现了诃子多成分、多靶点解草乌心脏毒性的作用特点。
6 分子对接
利多卡因(lidocaine)作为一种广谱抗心律失常药, 临床上常被运用于治疗乌头类药材引起的心律不齐室性心动过速[12, 13], 因此采用利多卡因作为分子对接的阳性对照药。以“lidocaine”作为关键词于GeneCards平台获得其相关靶点基因, 通过relevance score进行排序, 最终获得排名靠前的10个靶点(表 4)。通过与40个诃子解草乌毒的潜在靶点进行比较, 发现Na+离子通道蛋白SCN5A (PUB ID: 4DCK)和凝血酶F2 (PUB ID: 1A2C)为共同靶点, 因此把其作为分子对接的候选靶点。在进行分子对接时, 补充两个感兴趣的分子对接候选靶点: K+离子通道蛋白KCNA3 (PUB ID: 4BGC)和Na+/Cl-神经递质转运蛋白SLC6A2 (PUB ID: 5I6X)。因此, 将SCN5A、F2、SLC6A2、KCNA3的PUB ID、诃子15个活性化合物和lidocaine的2D结构上传至SystemsDock, 获得docking scores (pKd/pKi), 发现诃子15个活性化合物与4个候选靶点(SCN5A、F2、SLC6A2、KCNA3)对接活性均好于阳性对照药利多卡因(图 3)。
讨论
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蒙医临床上除了外用生品草乌以外主要以炮制品入药, 即用诃子汤炮制或以诃子配伍使用。诃子为使君子科植物诃子(Terminalia chebula Retz.)或绒毛诃子(Terminalia chebula Retz, var. tomentella Kurt.)的干燥成熟果实, 蒙文名阿如拉, 味涩性平, 蒙医称其为“蒙药之王”, 具有祛变形三根所引起的诸疾、调理体素、解毒之功效且功效齐全。早在公元前200多年前的《月王药帝》就有蒙医对草乌与诃子搭配使用的论述, 在8世纪的《四部医典》中指出“黑黄永病的治疗中加诃子、草乌为宜”, 19世纪的《蒙药正典》也指出在含有草乌的方剂中必须搭配诃子[14]。
草乌的主要毒效成分为二萜类生物碱(乌头碱、次乌头碱和中乌头碱等), 心律失常是其主要心脏毒性表现且机制已明确[15]。关于诃子“解草乌毒”的现代研究颇多, 有学者[16]发现诃子中的鞣质成分能够保护草乌当中的生物碱类成分被水解同时诃子汤的酸性环境又可以促进草乌当中双酯型二萜类生物碱的水解。这与课题组前期认为的诃子中大量的鞣质能够与草乌当中的生物碱生成难溶性盐, 使得草乌当中的毒性成分二萜类生物碱相对强度下降, 在体内减缓吸收进而缓和其毒性[17, 18]相一致。课题组前期还发现诃子汤能够缓解乌头碱所致的心律失常[19], 同时也能降低草乌所造成的心肌毒性[20]。因此, 在化合物筛选过程中, 课题组补充了含量较高的鞣质成分作为活性成分进一步筛选, 最终从54个记载的化合物中获得7个鞣质、1个四环三萜、4个五环三萜、3个生物碱。
成分-靶点-通路网络显示, 诃子多成分、多靶点、多通路缓解草乌造成的心律不齐和心脏毒性的作用特点。而作用靶点的GO注释分析和靶点功能分析结果表明, 诃子解草乌毒主要与配体受体结合、肌肉收缩、循环系统进程、细胞内外离子交换等生物过程相关, 涉及细胞膜、神经元突触、离子通道、肌原纤维等细胞组分, G蛋白偶联受体、氧化还原酶、转录因子、转运通道等多种物质参与了离子跨膜转运、神经递质受体活性、离子通道活性等分子过程, 整个过程复杂多样。而靶点的通路分析结果显示, 诃子解草乌心律不齐作用主要涉及神经活性配体受体相互作用、钙离子信号通路、心肌细胞肾上腺素能信号通路、cAMP信号通路、雌激素信号通路等。
有研究认为, 草乌当中的二萜类生物碱抑制心肌细胞膜电压门控Na+通道失活, 使Na+持续内流而延长细胞膜除极化时程, 从而导致不同类型的心律失常[21, 22]。离子通道是一类调控细胞膜上Na+、K+、Ca2+、Cl-等离子跨膜交换和流动的功能蛋白质。而离子通道病变参与了大部分的心律不齐, 也是心源性猝死的主要原因[23, 24]。诃子潜在靶点结果显示, 诃子的潜在靶点包括钾离子电压门控性通道类(KCNA1、KCNA4、KCNA3、KCNA5、KCNH2)、Na+通道蛋白类(SCN5A)和Na+/Cl-神经递质转运蛋白(SLC6A2)。笔者选择了其中的KCNA3、KCNA3、SLC6A2分别和利多卡因、诃子的15个活性成分进行分子对接, 分子对接结果发现诃子15个活性成分与离子通道蛋白结合性优于阳性对照药利多卡因, 进一步验证了诃子治疗草乌引起的心律不齐靶点在离子通道的可靠性。同时, 分子对接提示与凝血酶F2的良好结合性也进一步为诃子治疗草乌引起的心律不齐等心脏毒性提供了新的思路。
诃子抗草乌引起的心脏毒性的KEGG通路中神经活性配体受体相互作用通路的关键蛋白与心脏功能密切相关[25, 26], 肾素-血管紧张素的上调是心血管功能恶化的关键[27];而钙离子信号通路与心肌细胞上的钙离子转运相关, 研究表明[28]钙离子拮抗剂减少钙离子超载, 抑制细胞外钙内流, 降低细胞内钙离子浓度, 降低心肌收缩力; 心肌细胞肾上腺素能信号通路中的β-肾上腺素受体的亚型同心脏的功能和心衰有着密切关系。心肌细胞的β受体能够激活腺苷酸环化酶, 进一步导致cAMP的升高, 同时, cAMP的靶点蛋白激酶A (PKA)能够磷酸化与心脏功能相关的重要蛋白(L型该离子通道、受磷蛋白等)[29]。第二信使cAMP是心功能重要调节器[30], 能够调节心脏收缩和舒张的强度和频率。因此, 根据诃子解草乌心脏毒性的靶点通路推测, 诃子能够多途径地调节心肌细胞的收缩和舒张, 进一步保护心脏的正常功能, 减轻草乌造成的心律不齐等心脏毒性, 但是现有研究少有对于诃子解草乌毒性的分子研究, 因此, 诃子对草乌造成的心律不齐的机制需要后期的实验验证。
综上所述, 本文运用网络药理学的方法提示诃子的15个活性成分作用于40个靶点, 经过多种蛋白功能, 涉及了多种通路和生物进程, 进一步体现诃子多成分-多靶点-多途径缓解草乌造成的心脏毒性, 同时也为后续研究诃子能解草乌毒这一传统蒙医理论的现代机制研究提供了方向。
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  • 国家自然科学基金资助项目(81274192)
  • 国家自然科学基金资助项目(81360677)
  • 国家自然科学基金资助项目(81473797)
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2018年第53卷第10期
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doi: 10.16438/j.0513-4870.2018-0506
  • 接收时间：2018-05-29
  • 首发时间：2026-01-15
  • 出版时间：2018-10-12
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  • 收稿日期：2018-05-29
  • 修回日期：2018-07-04
基金
国家自然科学基金资助项目(81274192)
国家自然科学基金资助项目(81360677)
国家自然科学基金资助项目(81473797)
作者信息
    1.中国中医科学院医学实验中心, 北京 100700
    2.内蒙古医科大学蒙医药学院, 内蒙古 呼和浩特 010110
    3.内蒙古民族大学蒙医药学院, 内蒙古 通辽 028000
    4.中央民族大学药学院, 北京 100081
    5.中国中医科学院中医药研究发展中心, 北京 100700

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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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