Exploration of Material Basis of "Clearing Excessive Fire of Liver-gallbladder" of Bile-processed Coptidis Rhizoma Based on UPLC-Q-TOF/MS Metabolomics and Molecular Docking Technology

DONG Zhaowei; TIAN Maoying; YANG Yu; WANG Jin; HUANG Qinwan

doi:10.13422/j.cnki.syfjx.20230748

您当前的位置：

首页 >

文章列表页 >

Exploration of Material Basis of "Clearing Excessive Fire of Liver-gallbladder" of Bile-processed Coptidis Rhizoma Based on UPLC-Q-TOF/MS Metabolomics and Molecular Docking Technology

更新时间：2023-11-07

- Exploration of Material Basis of "Clearing Excessive Fire of Liver-gallbladder" of Bile-processed Coptidis Rhizoma Based on UPLC-Q-TOF/MS Metabolomics and Molecular Docking Technology
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 23, Pages: 140-149(2023)
- 作者机构：
  
  成都中医药大学西南特色中药资源国家重点实验室，药学院，民族医药学院，成都 611137
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20230748
  CLC： R22;R28;R943.1;O657
- Published：05 December 2023，
  
  Published Online：17 March 2023，
  
  Received：06 January 2023，
- 稿件说明：
扫描看全文
董兆威,田茂颖,杨宇等.基于UPLC-Q-TOF/MS代谢组学和分子对接技术探索胆黄连“清肝胆实火”的物质基础[J].中国实验方剂学杂志,2023,29(23):140-149.

DONG Zhaowei,TIAN Maoying,YANG Yu,et al.Exploration of Material Basis of "Clearing Excessive Fire of Liver-gallbladder" of Bile-processed Coptidis Rhizoma Based on UPLC-Q-TOF/MS Metabolomics and Molecular Docking Technology[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):140-149.
董兆威,田茂颖,杨宇等.基于UPLC-Q-TOF/MS代谢组学和分子对接技术探索胆黄连“清肝胆实火”的物质基础[J].中国实验方剂学杂志,2023,29(23):140-149. DOI： 10.13422/j.cnki.syfjx.20230748.

DONG Zhaowei,TIAN Maoying,YANG Yu,et al.Exploration of Material Basis of "Clearing Excessive Fire of Liver-gallbladder" of Bile-processed Coptidis Rhizoma Based on UPLC-Q-TOF/MS Metabolomics and Molecular Docking Technology[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(23):140-149. DOI： 10.13422/j.cnki.syfjx.20230748.

摘要

目的

采用超高效液相色谱-四极杆-飞行时间串联质谱法（UPLC-Q-TOF/MS）代谢组学及分子对接技术研究胆黄连清肝胆实火的物质基础。

方法

采用UPLC-Q-TOF/MS代谢组学技术分析黄连、清炒黄连和胆黄连的整体化学成分，流动相0.1%甲酸水溶液（A）-乙腈（B）梯度洗脱（0~2 min，5%B；2~20 min，5%~65%B；20~40 min，65%~10%B；40~45 min，10%B；45~46 min，10%~95%B；46~49 min，95%B），电喷雾离子源（ESI），正、负离子模式扫描，扫描范围

80~1 200。在此基础上，结合偏最小二乘法-判别分析（PLS-DA）和方差分析筛选不同黄连炮制品之间的差异化合物。利用网络药理学和分子对接技术验证差异化合物与肝胆实火证的关联程度。

结果

共鉴定了33个化学成分，包括2个酚酸类、5个结合型胆汁酸类、26个生物碱类。PLS-DA和方差分析确认差异化合物16个，其中11个生物碱、5个结合型胆汁酸。在京都基因与基因组百科全书（KEGG）数据库进行通路富集分析得到差异成分与“肝胆实火”相关的疾病通路8条，“成分-靶点-通路”网络分析得到关键靶点磷脂酰肌醇4，5-二磷酸3-激酶催化亚基

亚型（PIK3CA）；分子对接结果显示，11种生物碱成分与PIK3CA均具有良好的结合能力。

结论

猪胆汁在胆黄连炮制中具有独特性，可以促进小檗红碱、二氢白屈菜红碱等11种生物碱的生成和溶出。结合分子对接结果和文献结果可认为小檗红碱、二氢白屈菜红碱、牛磺猪去氧胆酸等16个差异化合物是胆黄连清肝胆实火的物质基础。

Abstract

Objective

To explore the material basis of bile-processed Coptidis Rhizoma clearing excessive fire of liver-gallbladder based on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF/MS） metabolomics and molecular docking.

Method

UPLC-Q-TOF/MS metabolomics was used to analyze the chemical constituents of Coptidis Rhizoma， water-processed Coptidis Rhizoma and bile-processed Coptidis Rhizoma. Chromatographic separation was achieved with 0.1% formic acid aqueous solution（A）-acetonitrile（B） as the mobile phase in gradient elution（0-2 min， 5%B； 2-20 min， 5%-65%B； 20-40 min， 65%-10%B； 40-45 min， 10%B； 45-46 min， 10%-95%B； 46-49 min， 95%B）， and electrospray ionization（ESI） was applied and operated in positive and negative ion modes， the acquisition range was

80-1 200. Based on this， partial least squares-discriminant analysis（PLS-DA） and variance analysis were used to screen the differential compounds among the three products of Coptidis Rhizoma. Network pharmacology and molecular docking were used to verify the degree of association between differential compounds and excessive fire of liver-gallbladder syndrome.

Result

A total of 33 chemical constituents were identified， including 2 phenolic acids， 5 binding bile acids and 26 alkaloids. And 16 differential compounds were identified by multivariate statistical analysis， including 11 alkaloids and 5 binding bile acids. Pathway enrichment analysis in the Kyoto Encyclopedia of Genes and Genomes（KEGG） database yielded 8 pathways related to excessive fire of liver-gallbladder， and the key protein phosphatidylinositol 4，5-bisphosphate 3-kinase catalytic subunit alpha isoform（PIK3CA） was obtained according to the "component-target-pathway" network analysis. Molecular docking results showed that 11 alkaloids had good binding ability with PIK3CA.

Conclusion

Porcine bile is unique in the processing of bile-processed Coptidis Rhizoma， which can promote the production and dissolution of 11 alkaloids， including berberine and dihydrochelerythrine. Based on the results of molecular docking and reported pharmacological experiments， it can be concluded that 16 different compounds such as berberine， dihydrochelerythrine and taurohyodeoxycholic acid are the material basis of bile-processed Coptidis Rhizoma.

关键词

胆黄连肝胆实火物质基础代谢组学分子对接技术网络药理学超高效液相色谱-四极杆-飞行时间串联质谱法（UPLC-Q-TOF/MS）

Keywords

bile-processed Coptidis Rhizomaexcessive fire of liver-gallbladdermaterial basismetabolomicsmolecular docking techniquenetwork pharmacologyultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF/MS）

references

国家药典委员会.中华人民共和国药典：一部［M］.北京：中国医药科技出版社，2020：303.

董兆威，田茂颖，赵毅萌，等.基于表里关联和多元统计分析的黄连药材分级研究［J］.世界中医药，2022，17（9）：1214-1219.

RAN Q，WANG J，WANG L，et al.Rhizoma Coptidis as a potential treatment agent for type 2 diabetes mellitus and the underlying mechanisms：A review［J］.Front Pharmacol，2019，10：805.

王静，吕佳，袁子民，等.猪胆汁炮制对黄连中生物碱类成分溶出的影响［J］.中国实验方剂学杂志，2016，22（5）：5-8.

YUAN Z M，CHEN Y，GAO H，et al.Comparative pharmacokinetic profiles of three protoberberine-type alkaloids from raw and bile-processed Rhizoma Coptidis in heat syndrome rats［J］.Pharmacogn Mag，2017，13（49）：51-57.

王静，陈悦，袁子民.胆黄连在实热证大鼠体内的整合药代动力学与药效学的相关性［J］.药学学报，2016，51（1）：127-131.

孙莹莹，钟凌云，龚千锋.基于大鼠舌体表面扫描电镜探讨黄连及其炮制品寒热药性［J］.中药药理与临床，2011，27（1）：54-55.

冉倩，楼冠华，曾海蓉，等.基于UPLC-Q-Orbitrap HRMS和网络药理学分析胆黄连的泻肝胆实火机制［J］.中国实验方剂学杂志，2020，26（13）：181-189.

岳文杰，洪翠云，赵峰，等.基于代谢组学分析两种萎凋方式对白茶萎凋过程代谢物与品质的影响［J］.亚热带植物科学，2021，50（6）：430-438.

张少平，李洲，练冬梅，等.基于代谢组学分析马齿苋根茎叶中4种重要次生代谢产物［J］.福建农业学报，2021，36（12）：1414-1421.

陈育青，王小平，余瑞铭，等.基于UPLC-Q-TOF-MS结合网络药理学的闽南炮制陈皮活性成分及潜在靶点研究［J］.中国中医药现代远程教育，2022，20（17）：140-143.

卫瑞，杨琳娇，秦雪梅，等.基于UPLC-Q-TOF-MS/MS和分子网络技术快速鉴定芦笋茎皮中的化学成分［J］.药学学报，2022，57（9）：2839-2850.

郭忠会，郑雪莹，梁洁，等.UPLC-Q-TOF/MS技术结合UNIFI平台快速鉴定鸡骨草化学成分［J］.中国药房，2022，33（23）：2852-2857，2863.

邓小芳，陈鸿，王爽，等.基于UPLC-QTOF-MS/MS和TCMIP v2.0辨识胆南星防治中风的质量标志物［J］.中国实验方剂学杂志，2022，28（12）：174-182.

陈正涛，杨燕，梁清芝，等.基于网络药理学和分子对接的丹参酮ⅡA治疗糖尿病肾病的机制研究［J］.现代药物与临床，2022，37（11）：2457-2464.

彭懿，左姿，卿志星，等.基于HPLC-Q-TOF/MS技术鉴定博落回叶中化学成分［J］.中南药学，2016，14（5）：465-470.

杨鹏，卿志星，向峰，等.HPLC-Q-TOF/MS法鉴定血水草中的异喹啉类生物碱［J］.中成药，2017，39（7）：1439-1443.

贾昌平，黄雪丽，李允，等.HPLC-DAD/ESI-Q-TOF-MS分析两面针中的生物碱［J］.中国中药杂志，2013，38（8）：1198-1202.

杨鹏，卿志星，左姿，等.HPLC-Q-TOF/MS鉴定白屈菜中异喹啉类生物碱［J］.中国现代中药，2017，19（2）：174-182.

赵利娟，高文雅，顾欣如，等.蒲地蓝消炎口服液化学成分鉴定及归属研究［J］.中国中药杂志，2019，44（8）：1573-1587.

CHEN M，WANG P L，LI T，et al.Comprehensive analysis of Huanglian Jiedu decoction：Revealing the presence of a self-assembled phytochemical complex in its naturally-occurring precipitate［J］.J Pharm Biomed Anal，2021，195：113820.

王征，CLIFFORD M N.LC-MSn比较分析三种传统中草药中绿原酸及其衍生物组分（英文）［J］.药学学报，2008，43（2）：185-190.

单国顺，赵启苗，潘多，等.基于UHPLC-Q-TOF MS/MS的胆南星中胆汁酸类成分的质谱裂解规律研究［J］.质谱学报，2020，41（4）：340-350.

冯志毅，张强，纪亮，等.甘草汁炮制前后栀子仁不同极性部位HPLC指纹图谱及其化学模式识别研究［J］.沈阳药科大学学报，2022，39（4）：405-413.

GUO Q，NIU W L，LI X J，et al.Study on hypoglycemic effect of the drug pair of Astragalus Radix and Dioscoreae Rhizoma in T2DM rats by network pharmacology and metabonomics［J］.Molecules，2019，24（22）：4050.

张道平，姚昆鹏，刘起立，等.基于生物信息学探讨肝癌相关细胞焦亡基因及潜在中药预测［J］.中草药，2022，53（23）：7467-7476.

章淑萍.慢性乙型肝炎生活情志及饮食的辨证施护［J］.辽宁中医杂志，2005，32（8）：846-847.

邹斌，方朝晖，哈团结.黄药消肿汤外敷联合针刺治疗毒性弥漫性甲状腺肿肝火亢盛证37例［J］.环球中医药，2022，15（8）：1453-1456.

孙瑞芳，刘石柱，李瑞东，等.化浊抗纤保肝汤调控PI3K/AKT/mTOR途径抑制肝纤维化的机制研究［J］.天津中医药，2022，39（7）：911-916.

张和韡，田甜，肖遥，等.从神经内分泌角度探讨肝主疏泄调畅情志的现代理论内涵［J］.环球中医药，2018，11（6）：850-853.

张蔓娜，万亿，田彬，等.乙型肝炎肝纤维化患者肝组织黏着斑激酶和整合素β1的表达及临床意义［J］.国际流行病学传染病学杂志，2021，48（5）：374-379.

周滔，张声生，陈誩.120例非酒精性脂肪肝单元证特点分析与健脾清肝方临床疗效［J］.中西医结合肝病杂志，2009，19（4）：209-210.

张小娟，蒋珍秀，邹蕴，等.高血压病中医证型与眼底血管病变、颈动脉粥样硬化、胰岛素水平的相关性分析［J］.中医临床研究，2019，11（16）：11-14.

LIU Y M，WANG M，LUO Y Q，et al.MiRNA-target network analysis identifies potential biomarkers for traditional Chinese medicine（TCM） syndrome development evaluation in hepatitis B caused liver cirrhosis［J］.Sci Rep，2017，7（1）：11054.

WU H W，WALDBAUER K，TANG L Y，et al.Influence of vinegar and wine processing on the alkaloid content and composition of the traditional Chinese medicine Corydalis Rhizoma（Yanhusuo）［J］.Molecules，2014，19（8）：11487-11504.

JIANG X，HUANG L F，WU L B，et al.UPLC-QTOF/MS analysis of alkaloids in traditional processed Coptis chinensis Franch［J］.Evid Based Complement Alternat Med，2012，2012：942384.

CHEN H Y，YE X L，CUI X L，et al.Cytotoxicity and antihyperglycemic effect of minor constituents from Rhizoma Coptis in HepG2 cells［J］.Fitoterapia，2012，83（1）：67-73.

YANG S，CAO S J，LI C Y，et al.Berberrubine，a main metabolite of berberine，alleviates non-alcoholic fatty liver disease via modulating glucose and lipid metabolism and restoring gut microbiota［J］.Front Pharmacol，2022，13：913378.

饶近秋，王凯，张婷婷，等.小檗红碱及其衍生物的研究进展［J］.中国药物化学杂志，2021，31（5）：369-377.

YANG X，MA L M，SHAO H W，et al.Riboflavin-promoted in situ photoactivation of dihydroalkaloid prodrugs for cancer therapy［J］.J Med Chem，2022，65（23）：15738-15748.

IWASAKI H，OKU H，TAKARA R，et al.The tumor specific cytotoxicity of dihydronitidine from Toddalia asiatica Lam［J］.Cancer Chemother Pharmacol，2006，58（4）：451-459.

IWASAKI H，OKABE T，TAKARA K，et al.Tumor-selective cytotoxicity of benzo［c］phenanthridine derivatives from Toddalia asiatica Lam［J］.Cancer Chemother Pharmacol，2010，65（4）：719-726.

WU Y R，MA Y B，ZHAO Y X，et al.Two new quaternary alkaloids and anti-hepatitis B virus active constituents from Corydalis saxicola［J］.Planta Med，2007，73（8）：787-791.

QI Y C，DUAN G Z，MAO W，et al.Taurochenodeoxycholic acid mediates cAMP-PKA-CREB signaling pathway［J］.Chin J Nat Med，2020，18（12）：898-906.

LI L，LIU C，MAO W，et al.Taurochenodeoxycholic acid inhibited AP-1 activation via stimulating glucocorticoid receptor［J］.Molecules，2019，24（24）：4513.

BAO L G，HAO D C，WANG X，et al.Transcriptome investigation of anti-inflammation and immuno-regulation mechanism of taurochenodeoxycholic acid［J］.BMC Pharmacol Toxicol，2021，22（1）：23.

RODA A，PIAZZA F，BARALDINI M，et al.Taurohyodeoxycholic acid protects against taurochenodeoxycholic acid-induced cholestasis in the rat［J］.Hepatology，1998，27（2）：520-525.

WANG W J，ZHAO J F，GUI W F，et al.Tauroursodeoxycholic acid inhibits intestinal inflammation and barrier disruption in mice with non-alcoholic fatty liver disease［J］.Br J Pharmacol，2018，175（3）：469-484.

XIAO L X，QI L，ZHANG X L，et al.Liver injury in septic mice were suppressed by a camptothecin-bile acid conjugate via inhibiting NF-κB signaling pathway［J］.Life Sci，2020，257：118130.

BHARGAVA P，SMITH M D，MISCHE L，et al.Bile acid metabolism is altered in multiple sclerosis and supplementation ameliorates neuroinflammation［J］.J Clin Invest，2020，130（7）：3467-3482.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Bioinformatics Reveals Mechanism of Zuoguiwan in Treating Polycystic Ovary Syndrome

Metabolomics-based Analysis of Toxicity Changes of Aconiti Lateralis Radix Praeparata-Zingiberis Rhizoma in Sinitang Before and After Compatibility

Exploring Mechanism of Renshen Guben Oral Liquids in Treating Renal Fibrosis Based on Metabolomics and Network Pharmacology

Effect of Guilu Erxiangao on Alzheimer's Disease and Its Mechanism Based on Kidney-brain Correlation

Analysis of Material Basis and Mechanism of Sangjiang Ganmao Injection in Treatment of Common Cold Based on UPLC-Q-Orbitrap HRMS and Network Pharmacology

Related Author

ZHANG Jinrong

LI Haotian

HUANG Hongming

DENG Ali

ZHAO Min

PENG Wei

LIU Dongmei

FU Shu

Related Institution

School of Basic Medical Sciences，Hubei University of Chinese Medicine

Affiliated Hospital of Hubei University of Chinese Medicine

School of Pharmacy，Chengdu University of Traditional Chinese Medicine

Sichuan Province Orthopedic Hospital

Zigong First People's Hospital

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰