红参中原人参三醇型皂苷组在肠道菌群中体外转化及对肠道菌群的作用

doi:10.19894/j.issn.1000-0518.210031

摘要/Abstract

摘要： 采用高分离度快速液相色谱-四极杆-飞行时间质谱(RRLC-Q-TOF MS)和超高效液相色谱-三重四极杆质谱联用技术(UPLC-QQQ MS)对红参中原人参三醇型皂苷组在人肠道菌群中体外代谢转化产物进行定性定量分析,并应用16S rDNA高通量测序技术分析原人参三醇型皂苷组对人肠道菌群多样性的影响。 RRLC-Q-TOF MS采用Supelco Ascentis Express C₁₈色谱柱(50 mm×3.0 mm×2.7 μm),UPLC-QQQ-MS/MS采用Thermo Syncronis-C₁₈色谱柱(100 mm×2.1 mm×1.7 μm),均以0.1%甲酸水-乙腈为流动相进行梯度洗脱,质谱分析均采用电喷雾负离子模式采集。原人参三醇型皂苷组中的人参皂苷Re、Rg1在人肠道菌群孵育后,RRLC-Q-TOF MS通过对照品比对及高分辨质谱数据解析分析转化产物为Rg2、Rh1、F1和PPT。 UPLC-QQQ MS定量分析代谢产物的转化率,在代谢转化第60 h时,产物Rg2、Rh1、F1、PPT的生成率为6%、12%、11%和56%。应用Illumina Hiseq 2500平台对粪便样本肠道菌群进行基因测序,与空白组粪便比较,原人参三醇型皂苷组在门水平上显著增加Firmicutes相对丰度,显著降低Bacteroidetes、Proteobacteria相对丰度;在属水平上Prevotella_9、Faecalibacterium、Dialister相对丰度显著增加,显著降低Escherichia-Shigella、Dorea、Lachnoclostridium相对丰度。该研究为基于肠道菌群对原人参三醇型皂苷药效物质基础的确定和药效作用靶点提供依据。

关键词: 原人参三醇型皂苷组, 人肠道菌群, 代谢转化, 液相色谱-质谱联用, 微生物多样性

Abstract: In order to discover the metabolites of protopanaxatriol saponins from red ginseng in human intestinal flora via intestinal bacteria incubation model. The identification and quantification of protopanaxatriol saponins were determined by using rapid resolution liquid chromatography-quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF MS) and ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQ MS) , and applied 16S rDNA high-throughput sequencing technology to tinvestigate the effect of protopanaxatriol on intestinal microorganisms. The UPLC-Q-TOF MS analysis was performed on a Supelco Ascentis Express C₁₈ RRHD (50 mm×3.0 mm×2.7 μm) and a Thermo Syncronis-C₁₈RRHD(100 mm×2.1 mm×1.7 μm)was used to UPLC-QQQ MS. The gradient elution was column with acetonitrile and 0.1% formic acid water as mobile phase. The data was collected by electropray ionization in positive mode. The results show that the ginsenoside Re and Rg1 in the protopanaxatriol group, after incubated in the human intestinal flora, according to the contrast of the reference standards and accurate masses of molecules, the transformation products into Rg2, Rh1, F1 and PPT are identified by RRLC-Q-TOF MS. Through the quantitative analysis of the conversion rate of metabolites, at the period of 60 h incubation, the production rates of products Rg2, Rh1, F1 and PPT are about 6%, 12%, 11% and 56%, respectively. Illumina Hiseq 2500 platform was used to sequence intestinal flora of feces samples and compared with the feces of the blank group, the protopanaxatriol saponin group significantly increased the relative abundance of Firmicutes at the phylum level, significantly decreased Bacteroidetes, Proteobacteria, and significantly increased Prevotella_9, Faecalibacterium, Dialister at the genus level, and significantly decreased Escherichia-Shigella, Dorea, Lachnoclostridium. This study provides a basis for the determination of the pharmacodynamic material basis and pharmacodynamic targets of protopanaxatriol-type saponins based on the intestinal flora.

Key words: Protopanaxatriol, Human intestinal microflora, Metabolic transformation, Liquid chromatography-mass spectrometry, Micro-biodiversity

中图分类号:

O657.6

越皓, 周东月, 张美玉, 张琰, 戴雨霖, 郑飞, 朱英豪. 红参中原人参三醇型皂苷组在肠道菌群中体外转化及对肠道菌群的作用[J]. 应用化学, 2021, 38(3): 323-330.

YUE Hao, ZHOU Dong-Yue, ZHANG Mei-Yu, ZHANG Yan, DAI Yu-Lin, ZHENG Fei, ZHU Ying-Hao. In vitro Biotransformation of Protopanaxtriol Saponins from Red Ginseng by Intestinal Flora and Its Effect on Intestinal Flora[J]. Chinese Journal of Applied Chemistry, 2021, 38(3): 323-330.

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