花旗泽仁主要活性成分在小鼠体内肠道菌群中的代谢研究
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摘要
目的:通过对花旗泽仁主要活性成分及其代谢产物在体内肠道菌群中的代谢研究,探索花旗泽仁体内代谢过程,为确定花旗泽仁的药效物质及临床合理用药奠定基础。方法:成年ICR小鼠随机分为空白组(n=6)及给药组(n=12),给药组灌服花旗泽仁水煎液后,分别收集两组小鼠4 h和12 h累积粪便,动物处死后迅速打开腹腔,取出盲肠内容物。利用UPLC-Q-TOF-MS方法对粪便和盲肠内容物中各原型化合物及其代谢产物进行快速分离和含量测定。结果:花旗泽仁的主要活性成分人参皂苷Rb_1(G-Rb_1)在1 h达峰,泽泻醇A-24-醋酸酯(Alisol A24-acetate)和9-羟基-(10E,12E)-十八碳二烯酸(9-HODE)在给药后12 h含量达到最高峰,但12 h的盲肠内容物里却检测不到G-Rb_1,而Alisol A 24-acetate和9-HODE在48 h仍可被检出;G-Rb_1的代谢产物G-Rd在给药后4 h含量达到最高峰,8 h后G-F_2、C-K和Ppd含量相继达到较高水平,而Alisol A 24-acetate的代谢产物Alisol A含量在12 h达峰,且在48 h仍可检测到微量的Alisol A。结论:明确花旗泽仁中主要活性成分及其代谢产物在体内肠道菌群中的代谢规律,证实了肠道菌群对花旗泽仁的代谢转化具有重要作用。
Objective:To investigate the metabolism of the main active components and their metabolites of Huaqizeren in intestinal microflora in mice. Methods:Adult ICR mice were randomly divided into blank group(n=6)and administration group(n=12). After filling the decoction of Huaqizeren,two groups of mice 4 h and 12 h accumulated feces were collected,and the animals were quickly opened to the abdominal cavity and removed the content of the cecum. Faecal and cecal contents were rapidly separated and determined by UPLC-Q-TOF-MS analysis. Results:Ginsenoside Rb_1(G-Rb_1),the main active component of Huaqizeren,was at the peak of 1 h,and alisone A-24-acetate(alisol A 24-acetate)and9-hydroxyl-(10 E,12 E)-eighteen-carbon two enoic acid(9-HODE)reached the peak of the 12 h content after administration,but G-Rb_1 was not detected in the cecal contents for 12 h,while alisol A 24-acetate and 9-HODE could still be detected at 48 h. After the drug delivery,the content of G-Rd reached the highest peak,4 h and the content of G-F_2,C-K and Ppd reached a high level after 8 h,while the alisol A content of the alisol A 24-acetate metabolite was at the peak of 12 h and a trace amount of alisol A could be detected at 48 h. Conclusion:The metabolic regulation of the main active components and their metabolites in the intestinal flora of Huaqizeren confirmed that the intestinal flora plays an important role in the metabolic transformation of the Huaqizeren.
Objective:To investigate the metabolism of the main active components and their metabolites of Huaqizeren in intestinal microflora in mice. Methods:Adult ICR mice were randomly divided into blank group(n=6)and administration group(n=12). After filling the decoction of Huaqizeren,two groups of mice 4 h and 12 h accumulated feces were collected,and the animals were quickly opened to the abdominal cavity and removed the content of the cecum. Faecal and cecal contents were rapidly separated and determined by UPLC-Q-TOF-MS analysis. Results:Ginsenoside Rb_1(G-Rb_1),the main active component of Huaqizeren,was at the peak of 1 h,and alisone A-24-acetate(alisol A 24-acetate)and9-hydroxyl-(10 E,12 E)-eighteen-carbon two enoic acid(9-HODE)reached the peak of the 12 h content after administration,but G-Rb_1 was not detected in the cecal contents for 12 h,while alisol A 24-acetate and 9-HODE could still be detected at 48 h. After the drug delivery,the content of G-Rd reached the highest peak,4 h and the content of G-F_2,C-K and Ppd reached a high level after 8 h,while the alisol A content of the alisol A 24-acetate metabolite was at the peak of 12 h and a trace amount of alisol A could be detected at 48 h. Conclusion:The metabolic regulation of the main active components and their metabolites in the intestinal flora of Huaqizeren confirmed that the intestinal flora plays an important role in the metabolic transformation of the Huaqizeren.
引文
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[2]Yang C A,Liang C,Lin C L,et al. Impact of Enterobius vermicularis infection and mebendazole treatment on intestinal microbiota and host immune response[J]. Plos Neglected Tropical Diseases,2017,11(9):e0005963.
[3]YANG Xiuwei,HAO Meirong,Hattori Masao.Metabolite analys is for chemical const ituents of traditional Chinese medicines[M].Beijing:Chinese Medicinal and Pharm Science and Technology Press,2003:144-151.
[4]Zu X P,Lin Z,Xie H S,et al. Interaction of effective ingredients from traditional Chinese medicines with intestinal microbiota[J].China Journal of Chinese Materia Medica,2016(10):1766.
[5]Tao J H,Zhao M,Wang D G,et al. Biotransformation and metabolic profile of catalpol with human intestinal microflora by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2016(1010):163-169.
[6]Gao MX,Tang XY,Zhang FX. Biotransformation and metabolic profile of Xian-Ling-Gu-Bao capsule,a traditional Chinese medicine prescription,with rat intestinal microflora by ultraperformance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry analysis[J]. Biomed Chromatogr,2018,32(4):537.
[7]韩东卫,朱蕾,张宇驰,等.基于UPLC/QTOF-MS技术观察花旗泽仁对SD大鼠体内尿液代谢组学的影响[J].中医药信息,2018,35(1):51-54.
[8]Chen Y,Duan J A,Guo J,et al. Yuanhuapine-induced intestinal and hepatotoxicity were correlated with disturbance of amino acids,lipids,carbohydrate metabolism and gut microflora function:A rat urine metabonomic study[J]. Journal of Chromatography B,2016,1026:183-192.
[9]Yang J L,Hu Z F,Zhang T T,et al. Progress on the Studies of the Key Enzymes of Ginsenoside Biosynthesis[J]. Molecules,2018,23(3):589.
[10]Yan S,Wei P C,Chen Q,et al. Functional and structural characterization of aβ-glucosidase involved in saponin metabolism from intestinal bacteria[J]. Biochemical&BiophysicalResearchCommunications,2018,496(4):1349-1356.
[11]Dong W W,Zhao J,Zhong F L,et al. Biotransformation ofPanax ginsengextract by rat intestinal microflora:identification and quantification of metabolites using liquid chromatographytandem mass spectrometry[J]. Journal of Ginseng Research,2017,41(4):540-547.
[12]Liu C,Hu M,Guo H,et al. Combined Contribution of Increased Intestinal Permeability and Inhibited Deglycosylation of Ginsenoside Rb1 in Intestinal Tract to the Enhancement of Ginsenoside Rb1 Exposure in Diabetic Rats Following Oral Administration[J]. Drug Metabolism&Disposition,2015,43(11):1702-1710.
[13]Shin HY,Lee JH,Lee JY. Purification and characterization ofginsenosideRa-hydrolyzingβ-D-xylosidasefrom Bifidobacterium breve K-110 a Human Intestinal Anaerobic Bacterium[J]. Biol Pharm Bull,2003,26(8):1170-1173.
[14]Chen L,Zhou L,Huang J,et al. Single-and Multiple-Dose Trials to Determine the Pharmacokinetics,Safety,Tolerability,and Sex Effect of Oral Ginsenoside Compound K in Healthy Chinese[J]. Frontiers in Pharmacology,2018,8:965.
[15]Hou J,Xue J,Zhao X,et al. Octyl ester of ginsenoside compound K as novel anti-hepatoma compound:Synthesis and evaluation on murine H22 cells in vitro and in vivo[J]. Chemical Biology&Drug Design,2017,94(4):951.
[16]Huang Y,Liu H,Zhang Y,et al. Synthesis and Biological Evaluation of Ginsenoside Compound K Derivatives as a Novel Class of LXRαActivator[J]. Molecules,2017,22(7):1232.