柯仲成+杨楠+侯雪峰+王爱东+封亮+贾晓斌

[摘要]为研究肠道菌群对芍药苷在体内吸收和代谢的影响，该课题在离体培养的大鼠肠道菌群中，加入芍药苷进行厌氧孵育，分析芍药苷在48 h内的变化过程，探讨体外大鼠肠道菌群对芍药苷的代谢转化。采用UPLC监测芍药苷在不同孵育时间点的变化，色谱条件为：WelchromTM C₁₈色谱柱（4.6 mm×100 mm，5 μm），流动相0.1%甲酸（A）-乙腈（B），梯度洗脱，流速0.4 mL·min^-1，柱温30 ℃；同时运用UPLC-Q-TOF-MS/MS对转化产物进行分析鉴定（ESI离子源，正离子模式检测），结合化合物精确相对分子质量和MS裂解碎片信息，并辅以保留时间及文献数据对代谢产物进行结构解析，分析芍药苷肠道代谢规律。结果发现孵育24 h后，芍药苷已经完全被代谢转化，得到的代谢产物包括芍药内酯苷、芍药内酯苷元、脱酰基-芍药内酯苷、脱酰基-芍药内酯苷元和芍药内酯B等。通过代谢途径分析发现，离体的大鼠肠道菌群先将芍药苷转化成芍药内酯苷，然后通过脱葡萄糖、脱苯甲酰基和四元环裂解重排等多个途径被进一步代谢，将芍药苷逐步转化生成相对分子质量更小、疏水性更强的代谢产物，从而更好被肠道吸收。

[关键词]芍药苷； 肠道菌群； 生物转化

[Abstract]In order to clarify the effect of intestinal flora on the absorption and metabolism of paeoniflorin in vivo， the metabolism of paeoniflorin by rat intestinal flora was studied under the in vitro anaerobic condition. Paeoniflorin was incubated with rat anaerobic intestinal flora for 48 h， and UPLC was used to detect the changes of paeoniflorin at different incubation time points under the following chromatographic conditions：WelchromTM C₁₈ chromatographic column （4.6 mm×100 mm， 5 μm）， with 0.1% formic acid（A）-acetonitrile（B） as the mobile phase for gradient elution. The flow rate was 0.4 mL·min^-1， and column temperature was 30 ℃. UPLC-Q-TOF-MS with positive ion mode（ESI ion source） was applied to investigate the structural characterization of metabolic products. The structures of the metabolites were identified by accurate molecular weight， TOF-MS/MS fragmentation information， combined with retention time and literature data review， and the intestinal metabolic rules were then analyzed. After incubation for 24 h， the paeoniflorin was metabolized completely， and the resulting metabolites（albiflorin， albiflorinaglycone， deacylate albiflorin， deacylate albiflorin aglycone and paeonilactone-B） were detected in rat intestinal flora. The metabolic pathway analysis showed that the isolated rat intestinal flora first transformed peoniflorin into albiflorin， and then further metabolized by glucose removal， phenyl group removal， or four-membered ring pyrolysis and rearrangement. Paeoniflorin was gradually transformed into more hydrophobic metabolites with smaller molecular mass， which were better absorbed by the intestinal tract.

[Key words]paeoniflorin； intestinal bacteria； metabolism

doi：10.4268/cjcmm20162021

赤芍为毛茛科植物芍药Paeonia lactiflora Pall.或川赤芍P. veitchii Lynch.的干燥根，性苦，微寒，具有清热凉血、散瘀止痛、清泻肝火之功，可用于温毒发斑，血热吐衄，肝郁胁痛，经闭痛经，目赤肿痛，痈肿疮疡^[1]。赤芍总苷是其主要活性部位，而芍药苷则为代表性成分，研究表明其对心血管系统、神经系统均有显著保护作用，还有抗肿瘤、保肝、抗炎、抗氧化等药理活性^[2]。……