基于外翻肠囊模型考察人参、生姜单煎及合煎液纳米相态的肠吸收特性差异

Differences in Intestinal Absorption Characteristics of Nanophase in Single and Combined Decoctions of Ginseng Radix et Rhizoma and Zingiberis Rhizoma Recens Based on Everted Gut Sac Model

摘要:

目的 比较人参、生姜单煎和配伍合煎形成的纳米相态（NP）在大鼠肠吸收差异，探讨人参生姜合煎后形成的新NP对人参、生姜主要成分吸收的影响。方法 采用差速离心和透析技术分别富集人参、生姜单煎与合煎液中的NP；采用透射电子显微镜、粒度分析仪、纳米颗粒跟踪分析仪对其形态、粒径、Zeta电位、浓度等进行表征；采用外翻肠囊模型，以超高效液相色谱-三重四极杆串联质谱法（UPLC-QqQ-MS/MS）对人参、生姜单煎、合煎的NP肠吸收液中指标性成分进行分析，以单位面积实际累积吸收量（Q_实际）、吸收速率常数（K_a）、表观渗透系数（P_app）为评价指标，考察上述NP在十二指肠、空肠、回肠、结肠中的吸收特性。结果 人参、生姜单煎液与合煎液中均存在不规则球形NP，且合煎形成的NP中成分含量多低于单煎NP。其中，10种成分可吸收进入肠囊，主要吸收部位在小肠，P_app均>1×10^-5 cm·min^-1。与单煎液NP比较，人参皂苷Rb₁、人参皂苷Rf、4-姜酚、6-姜烯酚等成分的Q_实际及K_a在合煎液NP中显著升高，而人参皂苷Re、6-姜酚则显著降低（P<0.01）；除人参皂苷Re、人参皂苷Rd外，其余成分的P_app在合煎液NP中均显著升高（P<0.01）。此外，人参皂苷Rb₁、人参皂苷Rd、人参皂苷Ro的Q_实际最大肠段部位前移，人参皂苷Rg₁、人参皂苷Re、8-姜酚后移；人参皂苷Rb₁、人参皂苷Rg₁、人参皂苷Rd、人参皂苷Ro的K_a最大肠段部位及P_app最大肠段部位前移，人参皂苷Re、4-姜酚则后移。结论 人参、生姜合煎有助于促进二者有效成分的吸收，同时改变部分肠段对有效成分的吸收行为，该研究可为后续二药配伍机制研究提供参考。

Abstract:

Objective To compare the differences in intestinal absorption of nanophase（NP） formed by single decoction and combined decoction of Ginseng Radix et Rhizoma（GRR） and Zingiberis Rhizoma Recens（ZRR） in rats， and to investigate the effects of new NP formed by the combined decoction on the absorption of main components in GRR and ZRR.Method Differential centrifugation and dialysis techniques were used to enrich NP in the single and combined decoctions of GRR and ZRR， respectively. The microstructure， particle size， Zeta potential and concentration of the NP were analyzed by transmission electron microscope， particle size analyzer and nanoparticle tracking analyzer. Based on everted gut sac model， the index components in the intestinal absorption solution of NP from the single and combined decoctions were analyzed by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry（UPLC-QqQ-MS/MS）. The per unit area actual value of cumulative intestinal absorption（Q_actual）， absorption rate constant（K_a） and apparent permeability coefficient（P_app） were used as the evaluating indexes to investigate the absorption characteristics of the aforementioned NP in the duodenum， jejunum， ileum and colon.Result Irregularly spherical NP was present in the single and combined decoctions， and the contents of components in NP of the combined decoction were mostly lower than those in the single decoction. In these NP， ten components could be absorbed into the intestinal sac， with the main absorption site being the small intestine， and the P_app was greater than 1×10^-5 cm·min^-1. Compared with NP in the single decoction， the Q_actual and K_a of ginsenoside Rb₁， ginsenoside Rf， 4-gingerol and 6-shogaol were significantly increased in NP of the combined decoction， while ginsenoside Re and 6-gingerol were significantly decreased（P<0.01）. Except for ginsenoside Re and ginsenoside Rd， the P_app of the remaining constituents was significantly increased in NP of the combined decoction（P<0.01）. In addition， the maximum intestinal segment site of Q_actual was shifted forward for ginsenoside Rb₁， ginsenoside Rd and ginsenoside Ro， while shifted backward for ginsenoside Rg₁， ginsenoside Re and 8-gingerol. The maximal intestinal segment sites of K_a and P_app of ginsenoside Rb₁， ginsenoside Rg₁， ginsenoside Rd and ginsenoside Ro shifted forward， while ginsenoside Re and 4-gingerol were shifted backward.Conclusion The combined decoction of GRR and ZRR is helpful to promote the absorption of the effective components of the two， and changes the absorption behavior of the effective components in some intestinal segments. This study provides a reference for the subsequent research on the compatibility mechanism of the two medicines.