Effect of Gut Microbiota on the Metabolism of Chemical Constituents of Berberis kansuensis Extract Based on UHPLC-Orbitrap-MS Technique

Abstract

The dried stem bark of Berberis kansuensis is a commonly used Tibetan herbal medicine for the treatment of diabetes. Its main chemical components are alkaloids, such as berberine, magnoflorine and jatrorrhizine. However, the role of gut microbiota in the in vivo metabolism of these chemical components has not been fully elucidated. In this study, an ultra-high performance liquid chromatography method coupled with Orbitrap mass spectrometry (UHPLC-Orbitrap-MS) technology was applied to detect and identify prototype components and metabolites in rat intestinal contents and serum samples after oral administration of a B. kansuensis extract. A total of 16 prototype components and 40 metabolites were identified. The primary metabolic pathways of the chemical components from B. kansuensis extract were demethylation, desaturation, deglycosylation, reduction, hydroxylation, and other conjugation reactions including sulfation, glucuronidation, glycosidation, and methylation. By comparing the differences of metabolites between diabetic and pseudo-germ-free diabetic rats, we found that the metabolic transformation of some chemical components in B. kansuensis extract such as bufotenin, ferulic acid 4-O-β-D-glucopyranoside, magnoflorine, and 8-oxyberberine, was affected by the gut microbiota. The results revealed that the gut microbiota can affect the metabolic transformation of chemical constituents in B. kansuensis extract. These findings can enhance our understanding of the active ingredients of B. kansuensis extract and the key role of the gut microbiota on them.

Key words

Berberis kansuensis - Berberidaceae Gut microbiota - Metabolism - UHPLC-Orbitrap-MS - Traditional Tibetan medicine

Publication History

Received: 17 April 2021

Accepted after revision: 22 August 2021

Article published online:
14 September 2021

• References

• 1 Chinese Pharmacopoeia Commission. Drug Standards of Tibetan Medicines. Beijing: Ministry of Health of the Peopleʼs Republic of China; 1995
• 2 Yutuo YDGB. The Four Medical Tantras. Shanghai: Shanghai Scientific and Technical Publishers; 1987
• 3 Disi SJJC. Lan Liu Li. Shanghai: Shanghai Scientific and Technical Publishers; 2012
• 4 Li Y, Lv XM, Tang C, Lai XR, Zhang Y, Fan G. Quality evaluation of cortex berberidis from different geographical origins by simultaneous high performance liquid chromatography combined with statistical methods. Trop J Pharm Res 2016; 15: 1973-1981
• 5 Li Q, Du H, Wen HS, Xu T, Wang YJ, Lai XR, Fan G. Determination of six compounds in Berberidis Cortex and comparative study of its different species. China J Chin Mater Med 2019; 44: 968-974
• 6 Zhang Y, Meng XL, Yue LJ, Xiang L, Lai XR. Preliminary studies on Tibetan medicine Berberis Cortex on blood glucose level in diabetic mellitus mice. Prog Mod Biomed 2013; 13: 3619-3622
• 7 Fan G, Li Q, Xu XM, Du H, Xu T, Lai XR, Du LL. Quality evaluation of different Berberidis Cortex species based on ¹H-NMR metabolomics and anti-diabetic activity. China J Chin Mater Med 2020; 45: 4677-4685
• 8 Feng R, Shou JW, Zhao ZX, He CY, Ma C, Huang M, Fu J, Tan XS, Li XY, Wen BY. Transforming berberine into its intestine-absorbable form by the gut microbiota. Sci Rep 2015; 5: 12155
• 9 Akao T, Kida H, Kanaoka M, Hattori M, Kobashi K. Drug metabolism: intestinal bacterial hydrolysis is required for the appearance of compound K in rat plasma after oral administration of ginsenoside Rb1 from Panax ginseng. J Pharm Pharmacol 1998; 50: 1155-1160
• 10 Paek IB, Moon Y, Kim J, Ji HY, Kim SA, Sohn DH, Kim JB, Lee HS. Pharmacokinetics of a ginseng saponin metabolite compound K in rats. Biopharm Drug Dispos 2006; 27: 39-45
• 11 Zhou SS, Xu J, Zhu H, Wu J, Xu JD, Yan R, Li XY, Liu HH, Duan SM, Wang Z. Gut microbiota-involved mechanisms in enhancing systemic exposure of ginsenosides by coexisting polysaccharides in ginseng decoction. Sci Rep 2016; 6: 1-13
• 12 Swartz ME. UPLC^™: an introduction and review. J Liq Chromatogr Relat Technol 2005; 28: 1253-1263
• 13 Nováková L, Matysová L, Solich P. Advantages of application of UPLC in pharmaceutical analysis. Talanta 2006; 68: 908-918
• 14 Michalski A, Damoc E, Hauschild JP, Lange O, Wieghaus A, Makarov A, Nagaraj N, Cox J, Mann M, Horning S. Mass spectrometry-based proteomics using Q Exactive, a high-performance benchtop quadrupole Orbitrap mass spectrometer. Mol Cell Proteomics 2011; 10: M111.011015
• 15 Shi F, Guo C, Gong L, Li J, Dong P, Zhang J, Cui P, Jiang S, Zhao Y, Zeng S. Application of a high resolution benchtop quadrupole-Orbitrap mass spectrometry for the rapid screening, confirmation and quantification of illegal adulterated phosphodiesterase-5 inhibitors in herbal medicines and dietary supplements. J Chromatogr A 2014; 1344: 91-98
• 16 Thomas A, Geyer H, Schänzer W, Crone C, Kellmann M, Moehring T, Thevis M. Sensitive determination of prohibited drugs in dried blood spots (DBS) for doping controls by means of a benchtop quadrupole/Orbitrap mass spectrometer. Anal Bioanal Chem 2012; 403: 1279-1289
• 17 Liu Q, Qiu S, Yu H, Ke Y, Jin Y, Liang X. Selective separation of structure-related alkaloids in Rhizoma coptidis with “click” binaphthyl stationary phase and their structural elucidation with liquid chromatography-mass spectrometry. Analyst 2011; 136: 4357-4365
• 18 Deevanhxay P, Suzuki M, Maeshibu N, Li H, Tanaka K, Hirose S. Simultaneous characterization of quaternary alkaloids, 8-oxoprotoberberine alkaloids, and a steroid compound in Coscinium fenestratum by liquid chromatography hybrid ion trap time-of-flight mass spectrometry. J Pharm Biomed Anal 2009; 50: 413-425
• 19 Singh A, Bajpai V, Kumar S, Rawat AKS, Kumar B. Analysis of isoquinoline alkaloids from Mahonia leschenaultia and Mahonia napaulensis roots using UHPLC-Orbitrap-MSn and UHPLC-QqQLIT-MS/MS. J Pharm Anal 2017; 7: 77-86
• 20 Singh A, Bajpai V, Kumar S, Kumar B, Srivastava M, Arya KR, Sharma KR. Distribution and discrimination study of bioactive compounds from Berberis species using HPLC-ESI-QTOF-MS/MS with principle component analysis. Nat Prod Commun 2016; 11: 1807-1812
• 21 Singh A, Bajpai V, Srivastava M, Arya KR, Kumar B. Kumar, Rapid profiling and structural characterization of bioactive compounds and their distribution in different parts of Berberis petiolaris Wall. ex G. Don applying hyphenated mass spectrometric techniques. Rapid Commun Mass Spectrom 2014; 28: 2089-2100
• 22 Tian PP, Zhang XX, Wang HP, Li PL, Liu YX, Li SJ. Rapid Analysis of components in Coptis chinensis Franch by ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry. Pharmacogn Mag 2017; 13: 175
• 23 Xue BJ, Zhao YY, Miao Q, Miao PP, Yang XY, Sun GX, Su J, Ye J, Wei BH, Zhang YY. In vitro and in vivo identification of metabolites of magnoflorine by LC LTQ-Orbitrap MS and its potential pharmacokinetic interaction in Coptidis Rhizoma decoction in rat. Biomed Chromatogr 2015; 29: 1235-1248
• 24 Tian XT, Zhang YC, Li ZX, Hu P, Chen MC, Sun ZL, Lin YF, Pan GY, Huang CG. Systematic and comprehensive strategy for metabolite profiling in bioanalysis using software-assisted HPLC-Q-TOF: magnoflorine as an example. Anal Bioanal Chem 2016; 408: 2239-2254
• 25 Wang DX, Liu P, Cui J, Wang JL, Cheng GY. Studies on intestinal absorption of Kaixinsan and its components. Chin J Drug Appl Monit 2007; 3: 10-14
• 26 Ma JY, Feng R, Tan XS, Ma C, Shou JW, Fu J, Huang M, He CY, Chen SN, Zhao ZX. Excretion of berberine and its metabolites in oral administration in rats. J Pharm Sci 2013; 102: 4181-4192
• 27 Xu P, Xu C, Li XX, Li D, Li Y, Jiang JB, Yang P, Duan GL. Rapid Identification of berberine metabolites in rat plasma by UHPLC-Q-TOF-MS. Molecules 2019; 24: 1994
• 28 Xu J, Chen HB, Li SL. Understanding the molecular mechanisms of the interplay between herbal medicines and gut microbiota. Med Res Rev 2017; 37: 1140-1185
• 29 Feng WW, Ao H, Peng C, Yan D. Gut microbiota, a new frontier to understand traditional Chinese medicines. Pharmacol Res 2019; 142: 176-191
• 30 Du H, Xu XM, Xu T, Li Q, Zhao CC, Yi H, Fan G. Effects of gut microbiota on five absorbed components of Berberis kansuensis in rat serum by HPLC-QqQ-MS. China J Chin Mater Med 2020; 45: 418-424
• 31 Long J, Song J, Zhong L, Liao Y, Liu L, Li X. Palmatine: A review of its pharmacology, toxicity and pharmacokinetics. Biochimie 2019; 162: 176-184
• 32 Yang WW, She LP, Yu K, Yan S, Zhang XF, Tian XY, Ma SR, Zhang XW. Jatrorrhizine hydrochloride attenuates hyperlipidemia in a high-fat diet-induced obesity mouse model. Mol Med Rep 2016; 14: 3277-3284
• 33 Feng T, Du H, Chen HT, Xiao QY, He Y, Fan G. Comparative analysis of genetic and chemical differences between four Berberis herbs based on molecular phylogenetic and HPLC methods. Biol Pharm Bull 2018; 41: 1870-1873
• 34 Du H, Li Q, Yi H, Xu T, Xu XM, Kuang TT, Zhang J, Huang AQ, Fan G. Anti-diabetic effects of Berberis kansuensis extract on type 2 diabetic rats revealed by ¹H-NMR-based metabolomics and biochemistry analysis. Chem Biodivers 2020; 17: e2000413
• 35 Chao PC, Li YX, Chang CH, Shieh JP, Cheng JT, Cheng KC. Cheng, Investigation of insulin resistance in the popularly used four rat models of type-2 diabetes. Biomed Pharmacother 2018; 101: 155-161
• 36 Jimeno R, Brailey PM, Barral P. Quantitative polymerase chain reaction-based analyses of murine intestinal microbiota after oral antibiotic treatment. JoVE 2018; e58481
• 37 Zhao L, Huang YY, Lu L, Yang W, Huang T, Lin ZS, Lin CY, Kwan H, Wong HLX, Chen Y. Saturated long-chain fatty acid-producing bacteria contribute to enhanced colonic motility in rats. Microbiome 2018; 6: 107