A new strategy for pharmacodynamic substance screening and research on gut microbiota pathway mechanisms based on UPLC-Q-orbitrap-MS and 16S rRNA
)Abstract
To establish a progressive research strategy for “colonic components analysis - efficacy verification and mechanism exploration - gut microbiota”, screen pharmacodynamic substances, and investigate their mechanism via gut microbiota.
The pharmacodynamics of Gegen Qinlian decoction (GQD) were assessed using a mouse model of dextran sulfate sodium-induced ulcerative colitis (UC). Ultra-performance liquid chromatography-quadrupole-orbitrap mass spectrometer was used to identify the prototype and metabolic components of GQD in the colon during UC. To analyze the structure and function of characteristic genera of GQD and its active components, 16S rRNA sequencing was performed.
We identified 67 prototypic and 14 metabolic components of GQD in the UC colon. The primary prototype components are flavonoids and alkaloids, including puerarin (PUE), baicalin (BAI), and berberine (BER). The metabolism was predominantly sulfonation. Efficacy verification showed that the main active components, puerarin, baicalin, and berberine, had good therapeutic effects on UC. The results of 16S rRNA gene sequencing showed that GQD improved UC by regulating the structure and function of the gut microbiota. The abundance of gut microbiota involved in the metabolism of the prototype components was influenced by the corresponding components. The function prediction results showed that PUE was the most comparable to GQD, with 24 consistent pathways. BAI and BER showed comparable gut microbiota regulation pathways. Characteristic pathways of BER include glucometabolic processes.
This study focused on the key issues in the gut microbiota pathway and developed a progressive research strategy to understand the transformation mechanisms of colonic components. This research systematically analyzed the active components and metabolic transformation of GQD in the colon during the pathological state of UC, as well as changes in the structure and function of the gut microbiota, clarified the mechanism of GQD and its active components in improving UC via the gut microbiota pathway.
Keywords
References
Ordás I, Eckmann L, Talamini M, Baumgart DC, Sandborn WJ. Ulcerative colitis. Lancet. 2012;380(9853):1606-1619.
Li T, Li Y, Zhang CJ, et al. Study on serum pharmacodynamic material basis of Gegen Qinlian Decoction in treatment of ulcerative colitis based on UHPLC-Q-Orbitrap-MS. Zhongguo Zhongyao Zazhi. 2022;47(4):1073-1084.
Wei MQ, Li H, Li QF, et al. Based on network pharmacology to explore the molecular targets and mechanisms of Gegen Qinlian Decoction for the treatment of ulcerative colitis. BioMed Res Int. 2020;2020:5217405.
Li QM, Cui Y, Xu BC, et al. Main active components of Jiawei Gegen Qinlian decoction protects against ulcerative colitis under different dietary environments in a gut microbiota-dependent manner. Pharmacol Res. 2021;170:105694.
Li YL, Li N, Liu JJ, et al. Gegen Qinlian Decoction alleviates experimental colitis and concurrent lung inflammation by inhibiting the recruitment of inflammatory myeloid cells and restoring microbial balance. J Inflamm Res. 2022;15:1273-1291.
Wang X, Huang S, Zhang M, et al. Gegen Qinlian decoction activates AhR/IL-22 to repair intestinal barrier by modulating gut microbiota-related tryptophan metabolism in ulcerative colitis mice. J Ethnopharmacol. 2023;302(Pt B):115919.
Braune A, Blaut M. Deglycosylation of puerarin and other aromatic C-glucosides by a newly isolated human intestinal bacterium. Environ Microbiol. 2011;13(2):482-494.
Nakamura K, Zhu S, Komatsu K, Hattori M, Iwashima M. Deglycosylation of the isoflavone C-glucoside puerarin by a combination of two recombinant bacterial enzymes and 3-oxo-glucose. Appl Environ Microbiol. 2020;86(14): e00607-e00620.
Cheng H, Liu J, Tan YZ, Feng WW, Peng C. Interactions between gut microbiota and berberine, a necessary procedure to understand the mechanisms of berberine. J Pharm Anal. 2022;12(4):541-555.
Kang MJ, Ko GS, Oh DG, et al. Role of metabolism by intestinal microbiota in pharmacokinetics of oral baicalin. Arch Pharm Res (Seoul). 2014;37(3):371-378.
Akao T, Akao T, Kobashi K. Glycyrrhizin beta-D-glucuronidase of Eubacterium sp. from human intestinal flora. Chem Pharm Bull. 1987;35(2):705-710.
Akao T, Akao T, Kobashi K. Glycyrrhizin stimulates growth of Eubacterium sp. strain GLH, a human intestinal anaerobe. Appl Environ Microbiol. 1988;54(8):2027-2030.
Li T, Liu F, Zhang XY, et al. Compatibility mechanism of Gegen Qinlian Decoction in treatment of ulcerative colitis based on network pharmacology. Zhongguo Zhongyao Zazhi. 2022;47(13):3619-3628.
Xu XZ, Gao ZZ, Yang FQ, et al. Antidiabetic effects of Gegen Qinlian Decoction via the gut microbiota are attributable to its key ingredient berberine. Dev Reprod Biol. 2020;18(6):721-736.
Akao T, Hayashi T, Kobashi K, et al. Intestinal bacterial hydrolysis is indispensable to absorption of 18 beta-glycyrrhetic acid after oral administration of glycyrrhizin in rats. J Pharm Pharmacol. 1994;46(2):135-137.
Ma SR, Tong Q, Lin Y, et al. Berberine treats atherosclerosis via a vitamine-like effect down-regulating Choline-TMA-TMAO production pathway in gut microbiota. Signal Transduct Targeted Ther. 2022;7(1):207.
Wang Y, Tong Q, Ma SR, et al. Oral berberine improves brain dopa/dopamine levels to ameliorate Parkinson's disease by regulating gut microbiota. Signal Transduct Targeted Ther. 2021;6(1):77.
Choi EJ, Kim GH. The antioxidant activity of daidzein metabolites, O-desmethylangolensin and equol, in HepG2 cells. Mol Med Rep. 2014;9(1):328-332.
Sekikawa A, Wharton W, Butts B, et al. Potential protective mechanisms of S-equol, a metabolite of soy isoflavone by the gut microbiome, on cognitive decline and dementia. Int J Mol Sci. 2022;23(19):11921.
Jeon YD, Lee JH, Lee YM, Kim DK. Puerarin inhibits inflammation and oxidative stress in dextran sulfate sodium-induced colitis mice model. Biomed Pharmacother. 2020;124:109847.
Elmaksoud HAA, Motawea MH, Desoky AA, Elharrif MG, Ibrahimi A. Hydroxytyrosol alleviate intestinal inflammation, oxidative stress and apoptosis resulted in ulcerative colitis. Biomed Pharmacother. 2021;142:112073.
Zhang L, Lin G, Zuo Z. Involvement of UDP-glucuronosyltransferases in the extensive liver and intestinal first-pass metabolism of flavonoid baicalein. Pharm Res (N Y). 2007;24(1):81-89.
Zhang L, Lin G, Chang Q, Zuo Z. Role of intestinal first-pass metabolism of baicalein in its absorption process. Pharm Res (N Y). 2005;22(7):1050-1058.
Mroczyńska M, Galecka M, Szachta P, Kamoda D, Libudzisz Z, Roszak D. Beta-glucuronidase and Beta-glucosidase activity in stool specimens of children with inflammatory bowel disease. Pol J Microbiol. 2013;62(3):319-325.
Gao S, Sun RJ, Singh R, et al. The role of gut microbial β-glucuronidase in drug disposition and development. Drug Discov Today. 2022;27(10):103316.
Wu WJ, Yan R, Li T, Li YP, Zhou RN, Wang YT. Pharmacokinetic alterations of rhubarb anthraquinones in experimental colitis induced by dextran sulfate sodium in the rat. J Ethnopharmacol. 2017;198:600-607.
Laryushina Y, Samoilova-Bedych N, Turgunova L, et al. Alterations of the gut microbiome and TMAO levels in patients with ulcerative colitis. J Clin Med. 2024;13(19):5794.
Tikunov AY, Shvalov AN, Morozov VV, et al. Taxonomic composition and biodiversity of the gut microbiome from patients with irritable bowel syndrome, ulcerative colitis, and asthma. Vavilovskii Zhurnal Genet Selektsii. 2021;25(8):864-873.
Ma X, Hu QC, Jiang T, et al. Dehydroevodiamine alleviates ulcerative colitis by inhibiting the PI3K/AKT/NF-κB signaling pathway via targeting AKT1 and regulating gut microbes and serum metabolism. Molecules. 2024;29(17):4031.
Scanu M, Toto F, Petito V, et al. An integrative multi-omic analysis defines gut microbiota, mycobiota, and metabolic fingerprints in ulcerative colitis patients. Front Cell Infect Microbiol. 2024;14:1366192.
Ni YH, Zhang Y, Zheng LJ, et al. Bifidobacterium and Lactobacillus improve inflammatory bowel disease in zebrafish of different ages by regulating the intestinal mucosal barrier and microbiota. Life Sci. 2023;324:121699.
Hirano A, Umeno J, Okamoto Y, et al. Comparison of the microbial community structure between inflamed and non-inflamed sites in patients with ulcerative colitis. J Gastroenterol Hepatol. 2018;33:1590-1597.
Wade H, Pan KC, Duan QH, et al. Akkermansia muciniphila and its membrane protein ameliorates intestinal inflammatory stress and promotes epithelial wound healing via CREBH and miR-143/145. J Biomed Sci. 2023;30(1):38.
Koh A, De Vadder F, Kovatcheva-Datchary P, Bäckhed F. From dietary fiber to host physiology: short-chain fatty acids as key bacterial metabolites. Cell. 2016;165(6):1332-1345.
Parada Venegas D, De la Fuente MK, Landskron G, et al. Short chain fatty acids (SCFAs)-mediated gut epithelial and immune regulation and its relevance for inflammatory bowel diseases. Front Immunol. 2019;10:277.
Liu HF, Feng X, Wang DF, et al. Altered metabolome and microbiome features provide clues in predicting recurrence of ulcerative colitis. J Pharm Biomed Anal. 2024;239:115864.
Li MJ, Jin M, Zhao L, et al. Tumor-associated microbiota in colorectal cancer with vascular tumor thrombus and neural invasion and association with clinical prognosis. Acta Biochim Biophys Sin. 2024;56(3):366-378.
Xu D, Wu QY, Liu WY, Hu GN, Meng HH, Wang JS. Therapeutic efficacy and underlying mechanisms of gastrodia elata polysaccharides on dextran sulfate sodium-induced inflammatory bowel disease in mice: modulation of the gut microbiota and improvement of metabolic disorders. Int J Biol Macromol. 2023;248:125919.
Jones LA, Sun EW, Martin AM, Keating DJ. The ever-changing roles of serotonin. Int J Biochem Cell Biol. 2020;125:105776.
Wu H, Denna TH, Storkersen JN, Gerriets VA. Beyond a neurotransmitter: the role of serotonin in inflammation and immunity. Pharmacol Res. 2019;140:100-114.
Liu N, Sun SQ, Wang PJ, Sun YN, Hu QJ, Wang XY. The mechanism of secretion and metabolism of gut-derived 5-hydroxytryptamine. Int J Mol Sci. 2021;22(15):7931.
Sifroni KG, Damiani CR, Stoffel C, et al. Mitochondrial respiratory chain in the colonic mucosal of patients with ulcerative colitis. Mol Cell Biochem. 2010;342(1–2):111-115.
McCann JR, Rawls JF. Essential amino acid metabolites as chemical mediators of host-microbe interaction in the gut. Annu Rev Microbiol. 2023;77:479-497.
White PJ, Newgard CB. Branched-chain amino acids in disease. Science. 2019;363(6427):582-583.
Wei Z, Zhao S, Qin X, et al. Research progress in metabolic mechanism of branched chain amino acids. Chin Heart J. 2023;35(2):213-217 [Chinese].
Zheng CH, Zhang N, Chang YJ, et al. Effect of different components of Kui Jie Shuang enema prescription on oxidative stress and energy metabolism in rats with experimental ulcerative colitis. J Hebei Tradit Chin Med Pharmacol. 2018;33(3):1-6 [Chinese].
Lavelle A, Sokol H. Gut microbiota-derived metabolites as key actors in inflammatory bowel disease. Nat Rev Gastroenterol Hepatol. 2020;17(4):223-237.
京公网安备11010802044758号