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Article(id=1200394155970122364, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1200394147019477416, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2024-0027, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1704729600000, receivedDateStr=2024-01-09, revisedDate=1714060800000, revisedDateStr=2024-04-26, acceptedDate=null, acceptedDateStr=null, onlineDate=1764125869027, onlineDateStr=2025-11-26, pubDate=1720713600000, pubDateStr=2024-07-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1764125869027, onlineIssueDateStr=2025-11-26, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1764125869027, creator=13701087609, updateTime=1764125869027, updator=13701087609, issue=Issue{id=1200394147019477416, tenantId=1146029695717560320, journalId=1189982191388893191, year='2024', volume='59', issue='7', pageStart='1897', pageEnd='2182', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1764125866894, creator=13701087609, updateTime=1764225115484, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1200810425920115296, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1200394147019477416, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1200810425920115297, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1200394147019477416, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2126, endPage=2134, ext={EN=ArticleExt(id=1200394156460855955, articleId=1200394155970122364, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=Study on the material basis and molecular mechanism of Rhei Radix et Rhizoma-Persicae Semen combination in activating blood circulation and dispelling blood stasis based on efficacy experiments, network pharmacology and HPLC, columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

In this study, the effective substance group and molecular mechanism of Rhei Radix et Rhizoma-Persicae Semen combination (RRR-PS) in activating blood circulation and dispelling blood stasis were investigated by integrating efficacy experiments, network pharmacology and HPLC. The rat model of blood stasis syndrome was established, and the blood rheology index and coagulation four comprehensive evaluation were carried out. The results showed that compared with the model group, the whole blood viscosity, erythrocyte sedimentation rate and erythrocyte aggregation index of the rats in the RRR-PS group were significantly callback (P < 0.01). Network pharmacology found that RRR-PS combination exerted the effect of activating blood circulation and dispelling blood stasis by acting on calmodulin-1 (CALM1), nitric oxide synthase-2 (NOS2), glucocorticoid receptor (NR3C1) and other targets, regulating platelet activation, peroxisome proliferator-activated receptor (PPAR), vascular endothelial growth factor A (VEGF) and other signaling pathways, and found key components: sennoside B, (+)-catechin, emodin, physcion, rhein, aloe-emodin, chrysophanol, gallic acid. HPLC was used to explore the dissolution rate of key components. The results showed that the contents of catechin, emodin, chrysophanol and physcion were significantly increased after RRR-PS combination (P < 0.01). In summary, RRR-PS has a significant effect on promoting blood circulation and removing blood stasis, and its mechanism of action is related to promoting angiogenesis, anti-coagulation, anti-thrombosis and anti-inflammation. The efficacy is related to the change of solvent system and the large dissolution of catechin, emodin, chrysophanol and physcion after the RRR-PS combination. The results of the study can further provide a reference for the follow-up study on the active substances and mechanism of the RRR-PS combination. Animal experiments have been approved by the Experimental Animal Committee of Shaanxi University of Traditional Chinese Medicine (No. SUCMDL20210309002).

, correspAuthors=Xia SHEN, Fan PING, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2024 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=Lin ZHU, Ying LIU, Jie SHEN, Bo-rui LI, Ke-xin YUE, Xia SHEN, Fan PING), CN=ArticleExt(id=1200394158021137161, articleId=1200394155970122364, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=基于功效实验-网络药理学-HPLC探讨大黄-桃仁配伍活血化瘀的物质基础及分子作用机制, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

本研究整合功效实验、网络药理学和HPLC探讨大黄-桃仁药对(Rhei Radix et Rhizoma-Persicae Semen combination, RRR-PS) 配伍发挥活血化瘀作用的功效物质群及分子作用机制。建立大鼠血瘀证模型, 进行血液流变指标和凝血四项综合评价, 结果显示, 与模型组相比, RRR-PS组大鼠的全血黏度、血沉、红细胞聚集指数等指标均明显回调(P < 0.01)。网络药理学发现RRR-PS通过作用于钙调蛋白1 (CALM1)、一氧化氮合酶2 (NOS2)、糖皮质激素受体(NR3C1) 等靶点, 调控血小板活化、过氧化物酶体增殖物激活受体(PPAR)、血管内皮生长因子(VEGF) 等信号通路发挥活血化瘀的功效, 并发现关键成分: 番泻苷B、儿茶素、大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、没食子酸。运用HPLC探究关键成分的溶出率, 结果发现RRR-PS配伍后, 儿茶素、大黄素、大黄酚、大黄素甲醚4种主要功效物质含量均显著提高(P < 0.01)。综上所述, RRR-PS活血化瘀功效显著, 其作用机制与促进血管生成、抗凝血、抗血栓、抗炎有关; 功效发挥与RRR-PS配伍后, 溶剂系统改变, 儿茶素、大黄素、大黄酚、大黄素甲醚大量溶出相关。研究结果可进一步为后续RRR-PS配伍活血化瘀功效物质及作用机制研究提供参考。动物实验已获得陕西中医药大学实验动物委员会批准(批准号: SUCMDL20210309002)。

, correspAuthors=沈霞, 平凡, authorNote=null, correspAuthorsNote=
*沈霞, Tel: 86-29-38185165, E-mail: ;
平凡, Tel: 13572408262, E-mail:
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J Pract Med (实用医学杂志), 2017, 33: 492-495., articleTitle=null, refAbstract=null), Reference(id=1201106979428200946, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[23], rfOrder=22, authorNames=null, journalName=null, refType=null, unstructuredReference=Maciej MM, Bo N, Kristina NE, et al. Complement and coagulation: strangers or partners in crime? [J]. Trends Immunol, 2007, 28: 184-192., articleTitle=null, refAbstract=null), Reference(id=1201106979512087029, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[24], rfOrder=23, authorNames=null, journalName=null, refType=null, unstructuredReference=Ban WJ, Chen L. Effect of plasma elastin peptide on hemodynamics in septic shock rats and the intervention effect of sivelestat sodium [J]. Chin Rem Clin (中国药物与临床), 2010, 10: 407-409., articleTitle=null, refAbstract=null), Reference(id=1201106979621138936, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[25], rfOrder=24, authorNames=null, journalName=null, refType=null, unstructuredReference=Pang T, Xie Z, Chen Y, et al. Simultaneously determination of 7 anthraquinone derivatives in the herb-pair of Radix et Rhizoma with HPLC method [J]. Chin J New Drugs (中国新药杂志), 2017, 26: 1079-1088., articleTitle=null, refAbstract=null), Reference(id=1201106979738579450, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[26], rfOrder=25, authorNames=null, journalName=null, refType=null, unstructuredReference=Zhang CC, Liu HZ, Wang YL, et al. Simultaneous determination of sixteen analytes and its dissolution rules in drug pair of Rheum palmatum and Prunus davidiana by HPLC-Q-Exactive Orbitrap HRMS [J]. 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WBV: Whole blood viscosity; ESR: Erythrocyte sedimentation rate; EAI: Erythrocyte aggregation index; PT: Prothrombin time; APTT: Activated partial thromboplastin time; FIB: Plasma fibrinogen; TT: Thrombin time; RRR-PS (4) and RRR-PS (9): Extracts of rhubarb with low and high-doses. A-D: The influence of each group on WBV (200 s<sup>-1</sup>), WBV (30 s<sup>-1</sup>), WBV (5 s<sup>-1</sup>), WBV (1 s<sup>-1</sup>); E-J: The influence of each group on ESR, EAI, PT, APTT, FIB and TT. <i>n</i> = 8, <span class="mag-xml-inline-formula"><tex-math id="M1">$ \overline{x} $</tex-math></span>±<i>s</i>. <sup>##</sup><i>P</i> < 0.01 <i>vs</i> control group; <sup>**</sup><i>P</i> < 0.01 <i>vs</i> model group , figureFileSmall=cmkghsoqEiH0qHxCpI9Qqg==, figureFileBig=CTpkFuNEzbJ7QIRBaVtyaA==, tableContent=null), ArticleFig(id=1201106974399230355, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=Ae4FoShQD/RJvcKiVXc1RQ==, figureFileBig=OvPJJDcnWbSmbqjbwhwPGA==, tableContent=null), ArticleFig(id=1201106974470533526, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Figure 2, caption= (RRR-PS)-component-target network , figureFileSmall=Ae4FoShQD/RJvcKiVXc1RQ==, figureFileBig=OvPJJDcnWbSmbqjbwhwPGA==, tableContent=null), ArticleFig(id=1201106974546031000, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=1393Qt1ZYyN8V73sUjd5WA==, figureFileBig=QtrpVnsQD5ZKmL5q3QP1eA==, tableContent=null), ArticleFig(id=1201106974625722778, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Figure 3, caption= GO enrichment analysis diagram , figureFileSmall=1393Qt1ZYyN8V73sUjd5WA==, figureFileBig=QtrpVnsQD5ZKmL5q3QP1eA==, tableContent=null), ArticleFig(id=1201106974701220251, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=hD4/oys9AQ/lzk1b3rj1BA==, figureFileBig=TPzflKbC15ttS26fSxcXcw==, tableContent=null), ArticleFig(id=1201106974780912027, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Figure 4, caption= Chromatograms of mixed reference substances of eight components (A), <i>Rheum officinale</i> Baill. (B) and <i>Rheum officinale</i> Baill.-Persicae Semen (C). 1: Gallic acid; 2: (+)-Catechin; 3: Sennoside B; 4: Aloe-emodin; 5: Rhein; 6: Emodin; 7: Chrysophanol; 8: Physcion , figureFileSmall=hD4/oys9AQ/lzk1b3rj1BA==, figureFileBig=TPzflKbC15ttS26fSxcXcw==, tableContent=null), ArticleFig(id=1201106974848020893, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
GroupIDTermP-value
Group00WP: 4462Platelet-mediated interactions with vascular and circulating cells1.74×10-4
Group01KEGG: 05410Hypertrophic cardiomyopathy2.34×10-3
Group02R-HSA: 1592316Initial activation of prometalloproteinase 1 (proMMP1)2.23×10-5
Group03KEGG: 00910Nitrogen metabolism1.74×10-4
Group04KEGG: 04064NF-kappa B signaling pathway3.50×10-3
Group05WP: 1982Sterol regulatory element-binding proteins (SREBP) signalling1.34×10-3
Group06WP: 229Irinotecan pathway7.45×10-5
Group07WP: 247Small ligand G protein-coupled receptors (GPCRs)1.17×10-7
Group08WP: 4136Fibrin complement receptor 3 signaling pathway1.95×10-3
Group09WP: 98Prostaglandin synthesis and regulation1.08×10-11
Group10WP: 554Angiotensin converting enzyme (ACE) inhibitor pathway7.39×10-10
Group11KEGG: 04924Renin secretion5.01×10-6
Group12KEGG: 04611Platelet activation1.67×10-5
Group13KEGG: 05204Chemical carcinogenesis6.11×10-7
Group14KEGG: 04261Adrenergic signaling in cardiomyocytes1.80×10-6
Group15KEGG: 03320Peroxisome proliferator-activated receptor (PPAR) signaling pathway1.40×10-3
Group16KEGG: 04726Serotonergic synapse1.48×10-9
Group17KEGG: 00590Arachidonic acid metabolism4.39×10-11
Group18WP: 167Eicosanoid synthesis4.39×10-11
Group19KEGG: 04370Vascular endothelial growth factor A (VEGF) signaling pathway6.80×10-3
Group20KEGG: 05133Pertussis6.11×10-6
Group21WP: 15Selenium micronutrient network4.67×10-10
Group22KEGG: 04020Calcium signaling pathway9.80×10-11
Group23KEGG: 04728Dopaminergic synapse1.98×10-9
Group24KEGG: 04610Complement and coagulation cascades4.40×10-8
Group25R-HSA: 376419Formation of nuclear receptor-mediator of RNA polymerase II transcription subunit 1 (NR-MED1) coactivator complex8.42×10-12
Group26R-HSA: 2161899Arachidonic acid is epoxidated to 8, 9/11, 12/14, 15-epoxyeicosatrienoic acid (EET) by cytochrome P450 (CYP (5))1.02×10-9
Group27R-HSA: 1566962Elastin degradation by elastin-degrading extracellular proteinases1.10×10-7
), ArticleFig(id=1201106974915129760, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Table 1, caption=

KEGG analysis of main targets

, figureFileSmall=null, figureFileBig=null, tableContent=
GroupIDTermP-value
Group00WP: 4462Platelet-mediated interactions with vascular and circulating cells1.74×10-4
Group01KEGG: 05410Hypertrophic cardiomyopathy2.34×10-3
Group02R-HSA: 1592316Initial activation of prometalloproteinase 1 (proMMP1)2.23×10-5
Group03KEGG: 00910Nitrogen metabolism1.74×10-4
Group04KEGG: 04064NF-kappa B signaling pathway3.50×10-3
Group05WP: 1982Sterol regulatory element-binding proteins (SREBP) signalling1.34×10-3
Group06WP: 229Irinotecan pathway7.45×10-5
Group07WP: 247Small ligand G protein-coupled receptors (GPCRs)1.17×10-7
Group08WP: 4136Fibrin complement receptor 3 signaling pathway1.95×10-3
Group09WP: 98Prostaglandin synthesis and regulation1.08×10-11
Group10WP: 554Angiotensin converting enzyme (ACE) inhibitor pathway7.39×10-10
Group11KEGG: 04924Renin secretion5.01×10-6
Group12KEGG: 04611Platelet activation1.67×10-5
Group13KEGG: 05204Chemical carcinogenesis6.11×10-7
Group14KEGG: 04261Adrenergic signaling in cardiomyocytes1.80×10-6
Group15KEGG: 03320Peroxisome proliferator-activated receptor (PPAR) signaling pathway1.40×10-3
Group16KEGG: 04726Serotonergic synapse1.48×10-9
Group17KEGG: 00590Arachidonic acid metabolism4.39×10-11
Group18WP: 167Eicosanoid synthesis4.39×10-11
Group19KEGG: 04370Vascular endothelial growth factor A (VEGF) signaling pathway6.80×10-3
Group20KEGG: 05133Pertussis6.11×10-6
Group21WP: 15Selenium micronutrient network4.67×10-10
Group22KEGG: 04020Calcium signaling pathway9.80×10-11
Group23KEGG: 04728Dopaminergic synapse1.98×10-9
Group24KEGG: 04610Complement and coagulation cascades4.40×10-8
Group25R-HSA: 376419Formation of nuclear receptor-mediator of RNA polymerase II transcription subunit 1 (NR-MED1) coactivator complex8.42×10-12
Group26R-HSA: 2161899Arachidonic acid is epoxidated to 8, 9/11, 12/14, 15-epoxyeicosatrienoic acid (EET) by cytochrome P450 (CYP (5))1.02×10-9
Group27R-HSA: 1566962Elastin degradation by elastin-degrading extracellular proteinases1.10×10-7
), ArticleFig(id=1201106974986432929, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
AnalyteCalibration equationLinear range /ng·mL-1r2
Sennoside By = 1 205.10x - 4 072.128.2-282.00.999 9
(+)-Catechiny = 312.59x - 1 100.124.2-242.00.999 6
Emodiny = 3 800.10x - 11 971.025.0-250.00.999 9
Physciony = 695.51x - 3 521.825.0-250.00.999 8
Rheiny = 3 169.50x - 14 085.031.6-316.00.999 9
Aloe-emodiny = 5 934.00x - 16 847.025.0-250.00.999 9
Chrysophanoly = 6 934.50x + 6 380.322.6-226.00.999 9
Gallic acidy = 3 251.40x - 8 772.627.4-274.00.999 9
), ArticleFig(id=1201106975070319011, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Table 2, caption=

Regression equations and linear ranges of 8 constituents

, figureFileSmall=null, figureFileBig=null, tableContent=
AnalyteCalibration equationLinear range /ng·mL-1r2
Sennoside By = 1 205.10x - 4 072.128.2-282.00.999 9
(+)-Catechiny = 312.59x - 1 100.124.2-242.00.999 6
Emodiny = 3 800.10x - 11 971.025.0-250.00.999 9
Physciony = 695.51x - 3 521.825.0-250.00.999 8
Rheiny = 3 169.50x - 14 085.031.6-316.00.999 9
Aloe-emodiny = 5 934.00x - 16 847.025.0-250.00.999 9
Chrysophanoly = 6 934.50x + 6 380.322.6-226.00.999 9
Gallic acidy = 3 251.40x - 8 772.627.4-274.00.999 9
), ArticleFig(id=1201106975166788006, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
AnalyteSample recovery rateRSD value
Sennoside B96.10%1.66%
(+)-Catechin100.12%1.54%
Emodin96.34%0.36%
Physcion103.42%1.28%
Rhein102.36%1.93%
Aloe-emodin98.54%1.84%
Chrysophanol98.40%1.02%
Gallic acid97.36%1.36%
), ArticleFig(id=1201106975271645607, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Table 3, caption=

Sample recovery rate and RSD of 8 constituents

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AnalyteSample recovery rateRSD value
Sennoside B96.10%1.66%
(+)-Catechin100.12%1.54%
Emodin96.34%0.36%
Physcion103.42%1.28%
Rhein102.36%1.93%
Aloe-emodin98.54%1.84%
Chrysophanol98.40%1.02%
Gallic acid97.36%1.36%
), ArticleFig(id=1201106975363920296, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
AnalyteRRRRRR-PS
Sennoside B43.325 ± 2.12940.588 ± 1.936
(+)-Catechin84.503 ± 3.743166.864 ± 2.447**
Emodin1.424 ± 0.0616.827 ± 0.573**
Physcion13.011 ± 0.58796.139 ± 4.804**
Rhein9.108 ± 0.7895.390 ± 0.753**
Aloe-emodin10.081 ± 0.3958.820 ± 0.306*
Chrysophanol7.532 ± 0.27638.309 ± 4.340**
Gallic acid4.505 ± 0.2300.764 ± 0.773**
), ArticleFig(id=1201106975456194987, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1200394155970122364, language=CN, label=Table 4, caption=

Comparison of the contents of 8 components in medicinal Rheum officinale Baill. and Rheum officinale Baill.-Persicae Semen. n = 10, $ \overline{x} $ ± s. *P < 0.05, **P < 0.01 vs RRR group

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AnalyteRRRRRR-PS
Sennoside B43.325 ± 2.12940.588 ± 1.936
(+)-Catechin84.503 ± 3.743166.864 ± 2.447**
Emodin1.424 ± 0.0616.827 ± 0.573**
Physcion13.011 ± 0.58796.139 ± 4.804**
Rhein9.108 ± 0.7895.390 ± 0.753**
Aloe-emodin10.081 ± 0.3958.820 ± 0.306*
Chrysophanol7.532 ± 0.27638.309 ± 4.340**
Gallic acid4.505 ± 0.2300.764 ± 0.773**
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基于功效实验-网络药理学-HPLC探讨大黄-桃仁配伍活血化瘀的物质基础及分子作用机制
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朱琳 1, 2 , 刘莹 1, 2 , 申洁 1, 2 , 李柏睿 1 , 岳可心 1 , 沈霞 1, 2, * , 平凡 3, *
药学学报 | 研究论文 2024,59(7): 2126-2134
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药学学报 | 研究论文 2024, 59(7): 2126-2134
基于功效实验-网络药理学-HPLC探讨大黄-桃仁配伍活血化瘀的物质基础及分子作用机制
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朱琳1, 2, 刘莹1, 2, 申洁1, 2, 李柏睿1, 岳可心1, 沈霞1, 2, * , 平凡3, *
作者信息
  • 1.陕西中医药大学药学院, 陕西 西咸新区 712046
  • 2.陕西省秦岭中草药应用开发工程技术研究中心, 陕西 西咸新区 712046
  • 3.西安交通大学第一附属医院药学部, 陕西 西安 710061

通讯作者:

*沈霞, Tel: 86-29-38185165, E-mail: ;
平凡, Tel: 13572408262, E-mail:
Study on the material basis and molecular mechanism of Rhei Radix et Rhizoma-Persicae Semen combination in activating blood circulation and dispelling blood stasis based on efficacy experiments, network pharmacology and HPLC
Lin ZHU1, 2, Ying LIU1, 2, Jie SHEN1, 2, Bo-rui LI1, Ke-xin YUE1, Xia SHEN1, 2, * , Fan PING3, *
Affiliations
  • 1. College of Pharmacy, Shaanxi University of Traditional Chinese Medicine, Xixian New Area 712046, China
  • 2. Engineering Research Centre for Application and Development of Chinese Herbal Medicine in the Qinling Mountains of Shaanxi Province, Xixian New Area 712046, China
  • 3. Department of Pharmacy, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China
出版时间: 2024-07-12 doi: 10.16438/j.0513-4870.2024-0027
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摘要
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本研究整合功效实验、网络药理学和HPLC探讨大黄-桃仁药对(Rhei Radix et Rhizoma-Persicae Semen combination, RRR-PS) 配伍发挥活血化瘀作用的功效物质群及分子作用机制。建立大鼠血瘀证模型, 进行血液流变指标和凝血四项综合评价, 结果显示, 与模型组相比, RRR-PS组大鼠的全血黏度、血沉、红细胞聚集指数等指标均明显回调(P < 0.01)。网络药理学发现RRR-PS通过作用于钙调蛋白1 (CALM1)、一氧化氮合酶2 (NOS2)、糖皮质激素受体(NR3C1) 等靶点, 调控血小板活化、过氧化物酶体增殖物激活受体(PPAR)、血管内皮生长因子(VEGF) 等信号通路发挥活血化瘀的功效, 并发现关键成分: 番泻苷B、儿茶素、大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、没食子酸。运用HPLC探究关键成分的溶出率, 结果发现RRR-PS配伍后, 儿茶素、大黄素、大黄酚、大黄素甲醚4种主要功效物质含量均显著提高(P < 0.01)。综上所述, RRR-PS活血化瘀功效显著, 其作用机制与促进血管生成、抗凝血、抗血栓、抗炎有关; 功效发挥与RRR-PS配伍后, 溶剂系统改变, 儿茶素、大黄素、大黄酚、大黄素甲醚大量溶出相关。研究结果可进一步为后续RRR-PS配伍活血化瘀功效物质及作用机制研究提供参考。动物实验已获得陕西中医药大学实验动物委员会批准(批准号: SUCMDL20210309002)。

大黄-桃仁药对  /  血瘀证  /  活血化瘀  /  分子机制  /  中药功效物质

In this study, the effective substance group and molecular mechanism of Rhei Radix et Rhizoma-Persicae Semen combination (RRR-PS) in activating blood circulation and dispelling blood stasis were investigated by integrating efficacy experiments, network pharmacology and HPLC. The rat model of blood stasis syndrome was established, and the blood rheology index and coagulation four comprehensive evaluation were carried out. The results showed that compared with the model group, the whole blood viscosity, erythrocyte sedimentation rate and erythrocyte aggregation index of the rats in the RRR-PS group were significantly callback (P < 0.01). Network pharmacology found that RRR-PS combination exerted the effect of activating blood circulation and dispelling blood stasis by acting on calmodulin-1 (CALM1), nitric oxide synthase-2 (NOS2), glucocorticoid receptor (NR3C1) and other targets, regulating platelet activation, peroxisome proliferator-activated receptor (PPAR), vascular endothelial growth factor A (VEGF) and other signaling pathways, and found key components: sennoside B, (+)-catechin, emodin, physcion, rhein, aloe-emodin, chrysophanol, gallic acid. HPLC was used to explore the dissolution rate of key components. The results showed that the contents of catechin, emodin, chrysophanol and physcion were significantly increased after RRR-PS combination (P < 0.01). In summary, RRR-PS has a significant effect on promoting blood circulation and removing blood stasis, and its mechanism of action is related to promoting angiogenesis, anti-coagulation, anti-thrombosis and anti-inflammation. The efficacy is related to the change of solvent system and the large dissolution of catechin, emodin, chrysophanol and physcion after the RRR-PS combination. The results of the study can further provide a reference for the follow-up study on the active substances and mechanism of the RRR-PS combination. Animal experiments have been approved by the Experimental Animal Committee of Shaanxi University of Traditional Chinese Medicine (No. SUCMDL20210309002).

Rhei Radix et Rhizoma-Persicae Semen  /  blood stasis syndrome  /  activating blood circulation and dispelling blood  /  molecular mechanism  /  functional substance of Chinese medicine
朱琳, 刘莹, 申洁, 李柏睿, 岳可心, 沈霞, 平凡. 基于功效实验-网络药理学-HPLC探讨大黄-桃仁配伍活血化瘀的物质基础及分子作用机制. 药学学报, 2024 , 59 (7) : 2126 -2134 . DOI: 10.16438/j.0513-4870.2024-0027
Lin ZHU, Ying LIU, Jie SHEN, Bo-rui LI, Ke-xin YUE, Xia SHEN, Fan PING. Study on the material basis and molecular mechanism of Rhei Radix et Rhizoma-Persicae Semen combination in activating blood circulation and dispelling blood stasis based on efficacy experiments, network pharmacology and HPLC[J]. Acta Pharmaceutica Sinica, 2024 , 59 (7) : 2126 -2134 . DOI: 10.16438/j.0513-4870.2024-0027
正文
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大黄(Rhei Radix et Rhizoma, RRR) 苦寒, 善泄热破积, 亦能入血分, 逐瘀通经, 《血证论》中言其: “能推陈致新, 以损阳和阴, 凡属逆于血分之中, 致有血不和处, 大黄之性, 亦无不达。盖其药气最盛, 故能克而制之。”桃仁(Persicae Semen, PS) 苦、甘, 平, 善活血祛瘀, 润肠通便。《神农本草经》称桃仁: “主瘀血, 血闭。”中医认为大黄与桃仁, 一柔一刚, 大黄得桃仁相配伍, 则专入血分, 加强了其破血积、下瘀血日久凝滞之功。桃仁得大黄相配伍, 在活血的同时, 其破积之力也明显增强。
现代研究表明大黄具有活血止血、抗凝、抗炎等药理作用。桃仁影响血液流动的速率, 发挥其自身的活血化瘀作用[1]。课题组前期研究发现不同配比的大黄-桃仁药对(RRR-PS) 会对脂多糖所致蓄血症大鼠模型, 以及寒凝血瘀大鼠模型产生活血作用[2, 3]。然而由于中药的复杂属性, 目前对于RRR-PS中活血化瘀的成分-靶点的对应关系不清楚, 药效物质基础尚不明确。本课题组在中药配伍理论的指导下, 通过大鼠血瘀证模型观察血液流变指标和凝血四项指标验证RRR-PS活血化瘀功效。进而采用网络药理学分析其功效物质及分子作用机制, 并利用HPLC分析RRR-PS配伍后大黄功效物质群的溶出率变化, 阐明RRR-PS相须为用活血化瘀的分子机制。为后期进一步开展RRR-PS及其复方的药效作用机制研究及其质量标志物研究奠定理论依据。
材料与方法
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仪器    LG-R-80A全自动血液流变学仪、LG-PABER-1型血小板聚集凝血因子分析仪(北京世帝科学仪器公司); 仪表恒温水浴锅(上海树立仪器仪表有限公司); Waters 2695高效液相色谱仪(美国Waters公司); GB204型电子分析天平(北京赛多利斯仪器系统有限公司); TDL80-2B离心机(上海安亭科学仪器厂); KQ-200KED超声波清洗机(江苏昆山市超声仪器有限公司)。
药品与试剂    实验大黄采于陕西省镇巴县, 桃仁(去皮尖) 饮片购于陕西省中医药大学中药房, 经胡本祥教授鉴定后确定均为正品。对照品番泻苷B (批号: 315A022)、(+)-儿茶素(批号: 708G024)、大黄素(批号: 1205A027)、大黄素甲醚(批号: 718C022)、大黄酸(批号: 526B023)、芦荟大黄素(批号: 531B022)、大黄酚(批号: 523D025) 和没食子酸(批号: 725G024) 购自西安灏洋生物有限公司, 其质量分数均大于98%。色谱甲醇购自西安灏洋生物有限公司。盐酸肾上腺素注射液(批号: H42019700) 购自中国武汉市大药业有限公司; 肝素钠(批号: 55656c) 购自南京新百药业有限公司; 乙醇浓度为95%, 购自天津市天士力化学试剂有限公司。
实验动物    SPF级雄性的健康Sprague-Dawley (SD) 大鼠, 体质量(270 ± 20) g, 购自空军军医大学实验动物中心, 许可证号SCXK-(军) 2019-0007。分笼喂养于陕西中医药大学药学院中药药理实验室。动物实验已获得陕西中医药大学实验动物委员会批准(批准号: SUCMDL20210309002)。
药物制备    据前期课题组研究[3], RRR-PS (1∶1) 时活血化瘀效果最佳, 分别称取酒制大黄250 g、桃仁饮片共250 g。混合后依次加8、6倍量水, 各热回流提取1 h, 过滤后将冷却的两次水煎液合并, 并于70 ℃条件下的水浴锅中进行水浴加热, 直到浓缩至相当于生药的1 g·mL-1, 将浓缩的药物水煎液放置-4 ℃冷藏备用。
分组与给药    SD大鼠共32只, 并随机分为对照组、模型组、RRR-PS (1∶1) 低剂量组、RRR-PS (1∶1) 高剂量组, 每组各8只。给药组低、高剂量组分别给予总生药剂量的临床等效量的4倍(0.64 g·kg-1)、9倍(1.44 g·kg-1)。对照组、模型组分别灌胃给予等体积的生理盐水, 每日一次, 连续给药21天。
造模    在第20天时, 除对照组外, 模型组、RRR-PS (1∶1) 低剂量组、RRR-PS (1∶1) 高剂量组进行急性血瘀模型复制, 分别两次给予模型大鼠注射0.8 mg·kg-1的盐酸肾上腺素注射液, 两次间隔4 h。在第一次注射2 h后, 将造模大鼠放置于0~2 ℃冰水中长达10 min, 造成大鼠急性血瘀模型。第二次静脉注射后, 大鼠禁食不禁水, 于最后一次给药后12 h时注射10%水合氯醛麻醉后进行腹主动脉取血。
网络药理学分析    本研究通过TCMSP、TCMID、BATMAN-TCM数据库并结合文献[4], 设定参数OB ≥ 30%, DL ≥ 0.18, 查找核对RRR-PS活性成分, 建立RRR-PS小分子数据库。通过PharmMapper在线分析平台、TCMSP和BATMAN-TCM数据库建立RRR-PS靶点库。将RRR-PS所对应的靶点在TTD、CTD、DrugBank数据库进行校验, 得到大黄-桃仁活血化瘀相关的靶点。将其导入String数据库, 获得蛋白-蛋白相互作用(PPI) 关系, 通过Cytoscape构建(RRR-PS)-成分-靶点网络。用Cytoscape软件中的Network Analyzer插件进行拓扑分析。应用DAVID数据库进行KEGG和GO功能富集。通过Cytoscape软件中ClueGo和CluePedia插件对靶点进行通路分析, 根据参数Kappa Score Threshold为0.4进行了分组, 将Kappa Score Threshold > 0.4的两个条目分属于同一组。
色谱条件    色谱柱为武本C18 (250 mm × 4.6 mm, 5 μm) 色谱柱; 流动相为甲醇(A)-0.2%磷酸水(B), 进行梯度洗脱: 0~5 min, 5%~30% A; 5~8 min, 30%~42% A; 8~13 min, 42%~53% A; 13~18 min, 53%~68% A; 18~35 min, 68%~100% A; 35~37 min, 100% A。体积流量为1.0 mL·min-1; 柱温为30 ℃; 检测波长为260 nm; 进样量为10 μL。
对照品溶液的制备    分别精密称取番泻苷B、(+)-儿茶素、大黄素、大黄素甲醚、大黄酸、芦荟大黄素、大黄酚、没食子酸对照品, 于10 mL量瓶中, 甲醇溶解并稀释至10 mL, 得质量浓度分别为0.158、0.125、0.125、0.125、0.113、0.141、0.121、0.137 mg·mL-1的对照品储备液, 在4 ℃下保存备用
供试品溶液的制备    取干燥大黄样品, 精密称定0.1 g, 置于100 mL具塞锥形瓶中, 再精密加入4.5 mL甲醇, 称定质量[5, 6]。超声处理(功率500 W, 频率40 kHz) 30 min后, 冷却至室温。补足失重, 摇匀置离心机以10 000 r·min-1离心, 取上清液, 0.22 μm微孔滤膜滤过处理, 即得大黄供试品溶液。大黄-桃仁药对, 选取大黄0.1 g, 桃仁0.1 g, 构成比1∶1, 后续操作同大黄单味药供试品溶液制备方法。
方法学考察
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线性关系考察    精密吸取各对照品储备液2、4、6、8、10、12、15、18、20 μL进行测定, 并记录出峰时间和峰面积。浓度为横坐标, 峰面积为纵坐标。
精密度    精密吸取各混合对照品溶液, 连续进样6次, 记录峰面积并计算各成分RSD。
稳定性    精密吸取同一供试品溶液, 分别在制备后的0、2、4、8、16、24 h进行测定并记录待测成分的峰面积计算RSD。
重复性    精密称取大黄样品粉末6份, 制备供试品溶液, 再分别进样并记录待测成分的峰面积值。
加样回收率    精密称取大黄干燥粉末6份, 分别加入高、中、低3个质量浓度对照品溶液, 每一个质量浓度下各取2份, 按供试品溶液的制备项下操作, 分别进样, 记录待测成分的峰面积值, 并计算回收率。
含量测定    精密称定0.1 g的各待测样品, 平行制样3份, 按供试品溶液的制备项下操作, 并按色谱条件项下进行8种化学成分的测定。记录各成分峰面积, 并且计算各成分在样品中相对应的质量分数, 再用SPSS软件分析。
数据分析    使用IBM SPSS Statistics 21.0统计软件进行数据统计分析, 组间数据采用单因素方差分析(one-way ANOVA), 两组样本分析采用独立样本t检验, 分析数据以P < 0.05为差异有统计学意义。
结果
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1 RRR-PS对血瘀证大鼠血液状态的影响
测定大鼠全血黏度(whole blood viscosity, WBV)、血沉(erythrocyte sedimentation rate, ESR)、红细胞聚集指数(erythrocyte aggregation index, EAI) 血液流变指标和凝血四项: 凝血酶原时间(prothrombin time, PT)、活化部分凝血活酶时间(activated partial thromboplastin time, APTT)、纤维蛋白原数(plasma fibrinogen, FIB)、凝血酶时间(thrombin time, TT)。与对照组相比, 模型组大鼠WBV、ESR、EAI、FIB指标均显著升高, PT、APTT、TT指标均显著减少(P < 0.01), 说明大鼠已出现急性血瘀症状。与模型组相比, RRR-PS配伍不同剂量组大鼠的WBV、ESR、EAI、PT、APTT、FIB、TT指标均明显回调(P < 0.01), 证实RRR-PS配伍具有活血化瘀功效(图 1)。
2 RRR-PS活血化瘀的功效物质及分子机制分析
通过数据库和文献挖掘[7], 共得到RRR-PS化学成分167个, 大黄的有效成分14个, 桃仁有效成分10个, 其中山柰酚(kaempferol) 和儿茶素[(+)-catechin] 为RRR和PS的共有成分。RRR-PS活血化瘀靶点87个, 通过构建PPI网络和(RRR-PS)-成分-靶点网络(图 2), 得到以下结果: RRR-PS共有靶点14个, 非共有靶点中大黄有43个, 桃仁有30个。其中, 番泻苷B (sennoside B)、儿茶素、大黄素(emodin)、大黄素甲醚(physcion)、大黄酸(rhein)、芦荟大黄素(aloe-emodin)、大黄酚(chrysophanol)、没食子酸(gallic acid) 的节点度较大(degree > 10), 表明它们为其发挥活血化瘀功效的关键成分。番泻苷B、儿茶素、大黄素、大黄素甲醚、大黄酸、芦荟大黄素、大黄酚、没食子酸是大黄的主要活性成分, 提示RRR-PS配伍后, 对RRR的功效物质溶出影响更大, 本实验将对其进行更深层次的研究。
根据结果网络分析, degree > 13的靶点糖皮质激素受体(glucocorticoid receptor, NR3C1)、凝血因子Ⅹ (coagulation factor X, F10)、二肽基肽酶-4 (dipeptidyl peptidase 4, DPP4)、中性粒细胞弹性蛋白酶(neutrophil elastase, ELANE)、基质金属蛋白酶-9 (matrix metalloproteinase 9, MMP9)、一氧化氮合酶3 (nitric oxide synthase 3, NOS3)、一氧化氮合酶2 (NOS2)、凝血因子Ⅱ (F2)、过氧化物酶体增生激活受体γ (PPARG)、基质金属蛋白酶-3 (MMP3) 可能为RRR-PS发挥活血化瘀功效的关键靶点。分析发现, 关键靶点中参与促进血管生成的靶点有NOS2、NOS3、MMP3、MMP9; 参与抗凝血和抗血栓的靶点有F2、F10、ELANE; 参与抗炎的靶点有DPP4、NOS2、NR3C1、PPARG。RRR-PS发挥活血化瘀功效与促进血管生成、抗凝血、抗血栓、抗炎有关, 其中促进血管生成与活血相关, 抗凝血、抗血栓与化瘀相关, 而抗炎则贯穿了活血化瘀整个进程。
RRR-PS在发挥活血化瘀功效时也有着共有的成分和靶点, 包括共有的成分山柰酚和儿茶素, 共有的靶点雄激素受体(AR)、钙调蛋白1 (CALM1)、羧酸酯酶1 (CES1) 等, 这说明RRR-PS发挥活血化瘀功效时是两者协同增效的作用, 使活血化瘀的功效增强。基于此两者相使配伍, 刚柔并济, 可防止活血化瘀太过, 共奏活血化瘀之效。
KEGG结果显示RRR-PS参与的调控通路156条, 富集于28个分组, 见表 1。参与活血化瘀的主要调控6条通路, 包括血小板活化、过氧化物酶体增殖物激活受体(PPAR) 信号通路、血管内皮生长因子(VEGF) 信号通路、钙信号通路、补体和凝血级联反应、弹性蛋白降解。其中靶点NOS2映射到钙信号通路, NOS3映射到血小板活化、VEGF信号通路、钙信号通路, PPARG映射到PPAR信号通路, ELANE、MMP3、MMP9映射到通路弹性蛋白降解, 靶点F2映射到通路血小板活化、补体和凝血级联反应, F10映射到通路补体和凝血级联反应。
图 3的GO mapping图表明, 87个靶点均具有生物学活性。生物过程(BP) 图中可以看出其涉及炎症反应、血管生成、凝血、血小板激胞溶质钙离子浓度的正调节等进程; 分子功能(MF) 图中可以看出, 其涉及到类固醇激素受体活性、成纤维细胞生长脂肪酸结合因子结合、一氧化氮合酶活性、钙调蛋白结合等功能。一氧化氮合酶活性与NOS2和NOS3有关, 其与促进血管生成和抗炎有关, 钙调蛋白结合与CALM1有关, 其与促进血管生成有关; 细胞成分(CC) 图中可以看出, 其涉及胞外区、电压门控钙通道复合物、质膜等。通过以上分析, 可以得到RRR-PS的分子功能、生物进程、细胞组分涉及到了抗炎、抗凝血、血管生成与RRR-PS活血化瘀的功效密切相关。
3 RRR-PS对RRR发挥活血化瘀作用的功效物质影响
3.1 方法学考察
3.1.1 线性关系
各成分线性关系结果见表 2, 8种成分线性关系良好。
3.1.2 精密度
番泻苷B、儿茶素、大黄素、大黄素甲醚、大黄酸、芦荟大黄素、大黄酚、没食子酸RSD值分别为1.83%、1.53%、1.38%、1.01%、1.17%、1.06%、0.61%、1.36%, RSD值均小于2.00%, 结果表明仪器精密度良好。
3.1.3 稳定性
番泻苷B、儿茶素、大黄素、大黄素甲醚、大黄酸、芦荟大黄素、大黄酚、没食子酸RSD值分别为0.48%、1.42%、1.37%、1.64%、1.86%、0.81%、1.24%、0.60%, 结果表明供试品溶液在24 h内稳定。
3.1.4 重复性
番泻苷B、儿茶素、大黄素、大黄素甲醚、大黄酸、芦荟大黄素、大黄酚、没食子酸平均质量分数的RSD值分别为0.30%、0.66%、1.14%、1.01%、1.59%、0.50%、0.84%、0.51%, 结果表明该方法的重复性良好。
3.1.5 加样回收率
各成分的加样回收率在96.10%~103.42%之间, RSD值分别为1.66%、1.54%、0.36%、1.28%、1.93%、1.84%、1.02%、1.36%, 详细结果见表 3。
3.2 含量测定
图 4可看出8个成分色谱峰的分离度良好。表 4结果表明, RRR和RRR-PS中除番泻苷B外的7种成分的含量均有显著性差异(P < 0.05)。RRR-PS配伍组中的儿茶素、大黄素、大黄酚、大黄素甲醚这4种成分的含量, 较RRR组中的含量显著提高(P < 0.01), RRR-PS组的大黄酸、没食子酸、芦荟大黄素的含量较RRR组的含量显著降低(P < 0.05), 表明RRR-PS配伍后会影响大黄功效物质的溶出率。
讨论
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血瘀证是中医临床常见证候。《诸病源候论》中认为许多疾病的病机在于淤血阻滞; 《证治准绳》蓄血篇曰“百病由污血者多”, 指出瘀血致病广泛。现代研究表明血瘀证是多种疾病的发病基础, 如心脑血管疾病、消化系统疾病、代谢性疾病、呼吸系统疾病等[8, 9]。中医临床中多以和血法、活血法、破血法等治疗血瘀证, RRR-PS配伍以破积活血见长。应用现代血液流变学和凝血四项指标可以探讨血液本质和活血化瘀的疗效原理[10, 11]。本实验观察了RRR-PS对大鼠血瘀证的疗效。造模后大鼠与对照组相比, 模型组血液流变学指标均显著升高, 凝血四项中除FIB指标延长外，均显著减少, 表明大鼠造模成功; 与空白组相比, 低、高剂量的RRR-PS配伍都可以使血液流变学和凝血四项指标明显回调, 活血化瘀效果显著。
研究结果显示, 筛选出的RRR-PS活血化瘀的关键成分包括番泻苷B、儿茶素、大黄素、大黄酚、大黄酸、芦荟大黄素、大黄素甲醚、没食子酸等关键成分均具有药理活性。RRR的蒽醌类成分对脑缺血性卒中具有良好的神经保护作用, 其中大黄酸能降低缺血再灌注损伤大鼠caspase-9的表达, 从而减少梗死面积[12]。芦荟大黄素可以清除氧自由基, 同时对大鼠脑缺血具有保护作用[13]。大黄素甲醚对缺血、缺氧脑损伤具有神经保护作用[10]。大黄酚具有抑制血管收缩、促进血液凝固, 抑制动脉硬化的作用[14]。大黄素能抑制花生四烯酸诱导的血栓聚集, 能够有效抑制血栓的形成[15]。大黄鞣质类成分有可降低纤溶酶活性作用, 其能促进血小板黏附、聚集及血栓形成[16]。番泻苷和儿茶素对血栓形成具有抑制作用[17]。没食子酸可以阻止血小板中核苷三磷酸二磷酸水解酶(NTPDase) 和5′-核苷酸酶和腺苷脱氨酶(ADA) 活性的增加[18]。
通过虚拟筛选RRR-PS发挥活血化瘀功效的作用靶点及通路分析, 发现F10、ELANE、MMP9、NOS3、NOS2、F2、PPARG、MMP3等关键靶点作用于血小板活化、PPAR信号通路、VEGF信号通路、钙信号通路、补体和凝血级联反应、弹性蛋白降解。其中血小板活化在血栓形成、动脉硬化、血管生成等方面有重要作用[19]; PPAR信号通路可以调节促炎反应, 防止其过度激活[20]; VEGF信号通路对血管稳态具有重要意义, VEGF是发育和病理性血管生成所必需的[21]; 钙信号通路与新生血管的形成有密切的关系[22]; 研究表明炎症反应与补体和凝血级联反应之间的串扰可能会导致系统的脑血管病的发生[23]; 弹性蛋白降解过程中由弹性蛋白酶天然产生的弹性蛋白肽进入血流可以发挥远程作用[24]。综上, RRR-PS发挥活血化瘀的作用机制与促进血管生成、抗凝血、抗血栓、抗炎有关。
RRR-PS相须为用, 大黄得桃仁之佐, 引大黄之力专入血分以破血, 桃仁得大黄之助, 直入下焦以化瘀。一猛一缓, 一锐一钝, 相互促进, 破积化瘀功倍。药效物质基础的变化可能是RRR-PS配伍后破积化瘀效果功倍的原因。既往的研究表明, RRR-PS配伍后游离蒽醌增加, 结合蒽醌减少, 这与PS中β-葡萄糖苷酶与结合蒽醌发生水解反应有关[25, 26]。并且本研究通过HPLC分析, 进一步发现RRR-PS共煎煮后, RRR中儿茶素、大黄素、大黄酚、大黄素甲醚的含量, 较单用RRR时的含量增加。说明这四种成分可能是游离蒽醌中发挥活血化瘀作用的主要功效物质。RRR中儿茶素、大黄素、大黄酚、大黄素甲醚四种功效物质的大量溶出, 使活血化瘀功效增强, 可能与PS中含有丰富的油脂性成分, RRR-PS共煎煮后改变了溶剂系统的溶解度有关。
综上所述, 本研究通过急性血瘀模型验证了RRR-PS配伍使用发挥活血化瘀的作用。进而通过“中药-成分-靶点”网络的构建、蛋白-蛋白互作网络分析、GO分析、靶标通路分析等探究RRR-PS发挥活血化瘀的功效物质群; 通过HPLC研究RRR-PS配伍后大黄功效物质群的溶出率变化。从物质基础及分子机制方面阐明RRR-PS配伍使用的科学性, 为临床中药配伍使用的研究提供参考。
作者贡献: 朱琳参与实验设计、动物实验研究及数据分析, 并完成论文初稿撰写; 刘莹参与动物实验研究及数据整理; 申洁参与实验数据整理; 李柏睿与岳可心参与动物实验研究; 沈霞参与实验设计验证与核实及研究资金获取; 平凡参与实验设计, 研究项目管理与指导及论文审阅与修订。
利益冲突: 本研究内容无任何利益冲突。
基金
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  • 国家中医药管理局中医药科学技术研究专项课题(GZY-KJS-2021-005)
  • 陕西省自然科学基础研究计划项目(2022JM-538)
  • 陕西省中医药中青年科技骨干人才项目(11610000MB29656749/2020-00894)
  • 陕西省中医药管理局中医药“双链融合”中青年科研创新团队项目(2022-SLRH-LJ-003)
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参考文献 引证文献
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2024年第59卷第7期
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doi: 10.16438/j.0513-4870.2024-0027
  • 接收时间：2024-01-09
  • 首发时间：2025-11-26
  • 出版时间：2024-07-12
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  • 收稿日期：2024-01-09
  • 修回日期：2024-04-26
基金
国家中医药管理局中医药科学技术研究专项课题(GZY-KJS-2021-005)
陕西省自然科学基础研究计划项目(2022JM-538)
陕西省中医药中青年科技骨干人才项目(11610000MB29656749/2020-00894)
陕西省中医药管理局中医药“双链融合”中青年科研创新团队项目(2022-SLRH-LJ-003)
作者信息
    1.陕西中医药大学药学院, 陕西 西咸新区 712046
    2.陕西省秦岭中草药应用开发工程技术研究中心, 陕西 西咸新区 712046
    3.西安交通大学第一附属医院药学部, 陕西 西安 710061

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*沈霞, Tel: 86-29-38185165, E-mail: ;
平凡, Tel: 13572408262, E-mail:
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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
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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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