体外共孵育法研究大鼠肠道菌群对灯盏生脉胶囊主要成分的代谢规律

    Metabolic Profiling of Essential Components in Dengzhan Shengmai Capsule Exposed to Rats Intestinal Flora Using an in Vitro Co-incubation Model

    • 摘要:
      目的 明确灯盏生脉胶囊(Dengzhan Shengmai capsule,DZSM)提取物在大鼠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提取物与雄性SD大鼠肠道菌群共孵育,采用超高效液相色谱-四极杆-静电场轨道阱质谱仪,通过数据依赖性扫描(data-dependent acquisition,DDA)的全谱扫描与平行反应监测(parallel reaction monitoring,PRM)模式的靶向识别相结合的数据采集策略,对DZSM主要成分的代谢产物进行鉴定分析,在此基础上阐释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 大鼠肠道菌群对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 clarify the metabolic profiling of Dengzhan Shengmai capsule(DZSM) under the action of intestinal flora in rats.
      METHODS An in vitro co-incubation method was adopted, the 16 representative components of DZSM and the DZSM extract were co-incubated with intestinal flora isolated from male SD rats. Ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap mass spectrometry(UPLC-Q-Exactive-Orbitrap MS) was applied for identifying metabolites of components in DZSM. An integrated data acquisition strategy was employed, incorporating both full-scanning mode utilizing data-dependent acquisition(DDA) and targeted recognition mode with parallel reaction monitoring(PRM). Then the biotransformation rules of the main active components in DZSM under the action of intestinal flora were elucidated.
      RESULTS A total of 118 metabolites were generated from the 16 representative components through metabolic transformation by intestinal flora. Based on data analysis, the potential metabolic pathways of various components were inferred. Specifically, flavonoids underwent hydrolysis, isomerization, glycosylation and other reactions under the action of intestinal flora; caffeoyl esters mainly underwent hydrolysis, methylation and dehydroxylation; saponins were dominated by deglycosylation; lignans mainly underwent demethylation and hydroxylation.
      CONCLUSION  The intestinal flora of rats can transform the components of DZSM extract. The main active components of DZSM are mainly metabolized through hydrolysis, hydroxylation and other pathways under the action of intestinal flora.

       

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