体外共孵育法研究肠道菌群对灯盏生脉胶囊主要成分的代谢规律

Metabolic regularity of intestinal flora on main compoents of Dengzhan Shengmai formula by in vitro co-incubation method

关键词:  灯盏生脉方  肠道菌群  代谢规律  体外共孵育法  UPLC-Q-Exactive-Orbitrap MS

Key words:  Dengzhan Shengmai formula  gut microbiota  Metabolic regularity  co-incubation in vitro  UPLC-Q-Exactive-Orbitrap MS

摘要:

目的 明确灯盏生脉胶囊(DZSM)提取物在肠道菌群作用下的代谢转化规律。方法 采用体外共孵育方法,将DZSM中16个代表性成分以及DZSM提取物与大鼠肠道菌群共孵育,采用UPLC-Q-Exactive-Orbitrap MS分析,通过数据依赖性扫描(DDA)的全谱分析与平行反应监测(PRM)模式的靶向识别相结合,对16个代表性成分及DZSM提取物的代谢产物进行鉴定分析,在此基础上分析DZSM主要活性成分在肠道菌群作用下的生物转化规律。结果 16个代表性成分经肠道菌群代谢转化生成118个代谢产物,基于数据分析推测出各类成分可能的代谢途径。其中黄酮类化合物在肠道菌群作用下发生水解、异构化、糖基化等反应；咖啡酰酯类以水解、甲基化、去羟基化为主；皂苷类以去糖基化为主；木脂素类以去甲基化、羟基化为主。结论 肠道菌群对DZSM提取物成分有转化作用,其主要活性成分在肠道菌群代谢中以水解、羟基化等代谢途径为主。

Abstract:

OBJECTIVE To determine the metabolic transformation rule of Dengzhan Shengmai (DZSM) extract under the action of intestinal flora. METHODS The 16 standards of DZSM and their extracts were co-incubated with the intestinal flora of rats by in vitro co-incubation method. The metabolites of 16 Standard of DZSM were identified and analyzed by using UPLC-Q-Exactive-Orbitrap MS analysis technology. Based on these findings, the transformation of extracts of DZSM after co-incubation was analyzed. RESULTS The metabolism of 16 representative components produced 118 metabolites through intestinal flora. Based on the data and literature, the possible metabolic pathways of various components were deduced, the metabolism of flavonoids in intestinal flora is mainly hydrolysis, isomerization and glycosylation, while caffeoyl esters are mainly hydrolyzed, methylated and dehydroxylated, and saponins are mainly deglycosylation, the lignans were mainly demethylated and hydroxylated. After extract of DZSM was incubated with intestinal flora of rats, the peak area of caffeoyl esters with small molecular weight increased. The peak area of aglycone such as flavonoid aglycone and ginsenoside aglycone mostly increased, while the peak area of glycosides mostly decreased. CONCLUSION Intestinal flora can transform extract of DZSM components, and the main active components of DZSM are mainly metabolized by hydrolysis, hydroxylation, etc.