山豆根中主要成分在Caco-2细胞模型上的转运特性

Transport Characteristics of Main Ingredients from Sophorae Tonkinensis Radix et Rhizoma in Caco-2 Cell Mode

目的:考察山豆根水提物中主要成分在Caco-2细胞模型上的转运特性,为山豆根的体内吸收特性研究提供参考。方法:以Caco-2细胞模型研究山豆根中主要成分从给药端向吸收端的转运透过情况,采用UPLC-MS考察主要药效成分苦参碱和氧化苦参碱的吸收透过情况,流动相0.1%甲酸-乙腈梯度洗脱,流速0.4 m L·min-1。质谱条件采用正离子选择性离子进行检测,扫描测定氧化苦参碱(m/z±H)265.1,苦参碱(m/z±H)249.1,计算二者的表观渗透系数。结果:在给药120 min时,苦参碱、氧化苦参碱的表观渗透系数分别为(11.43±0.41)×10-6,(3.91±0.70)×10-6cm·s-1。山豆根水提物中可检测到20种成分,包括苦参碱在内的5个主要组分(P2,P4,P6,P7,P25)有较高的透过率,6种组分(P3,P5,P9,P11,P14,P27)具有中等透过率,包括氧化苦参碱在内的其他组分透过率较低或不能透过细胞模型。结论:山豆根的主要成分中约55%易透过细胞模型,其中苦参碱在体外模型中极易被吸收,而氧化苦参碱在体外模型中具有低透过率,提示苦参碱是山豆根吸收进入体内的主要有效成分之一。

Objective： To investigate transport characteristics of main ingredients from Sophorae Tonkinensis Radix et Rhizoma in Caco-2 cell model and provide a reference for further studies on its in vivo absorption characteristics. Method： Transport of main ingredients in Sophorae Tonkinensis Radix et Rhizoma from apical side to basolateral side was studied by Caco-2 cell monolayer model. Permeability of oxymatrine and matrine from these two sides was determined by UPLC-MS,mobile phase was 0. 1% formic acid-acitonitrile by gradient elution at flow rate of 0. 4 m L·min- 1. Mass spectrum was operated in positive mode with oxymatrine（ m / z ± H） 265. 1 and matrine（ m/z ± H） 249. 1,their apparent permeability coefficients（ Papp） were calculated. Result： At 120 min after dosing,Pappof matine was（ 11. 43 ± 0. 41） × 10- 6cm·s- 1,indicating that transmittance of matine was high;while Pappof oxymatine was（ 3. 91 ± 0. 70） × 10- 6cm·s- 1,indicating that transmittance of oxymatine was lower.Twenty constituents were detected in aqueous extract of Sophorae Tonkinensis Radix et Rhizoma,five constituents of them（ P2,P4,P6,P7,P25） including matrine had high levels of transmittance,six constituents（ P3,P5,P9,P11,P14,P27） had medium levels of transmittance,these other substances had low levels of transmittance or had no transmittance at all,which included oxymatrine. Conclusion： Fifty-five percent of main components are easily transported through momolayer. Among all components, matrine can be absorbed easily in vitro cell model,however,permeability of oxymatrine is poorer than matrine in vitro. This study suggests that matrine may be one of major components that was aborsopted in vivo of Sophorae Tonkinensis Radix et Rhizoma.