板蓝根提取液中有效成分的吸收转运特性分析

目的:研究板蓝根中有效成分在Caco-2细胞模型中的吸收转运特征。方法:建立Caco-2细胞摄取、转运模型,考察受试化合物对该细胞的安全性,以跨膜电阻值、碱性磷酸酶活性和荧光素钠通透性3个指标检验细胞模型,考察浓度、时间、温度、抑制剂及p H对受试化合物吸收的影响。采用HPLC检测受试化合物,计算其表观渗透系数(Papp)与外排率。结果:板蓝根样品中精氨酸、腺苷能被小肠吸收细胞较好吸收,精氨酸Papp在1.01×10-6~1.35×10~(-6)cm·s~(-1),腺苷Papp在0.40×10~(-6)~0.71×10~(-6)cm·s~(-1)。2种成分在Caco-2细胞中膜通透性良好;精氨酸被吸收程度优于腺苷,精氨酸吸收率50%。板蓝根提取液中精氨酸、腺苷成分外排率(ER)均在1.0~2.0,存在P-糖蛋白(P-gp)外排转运蛋白介导两者的吸收转运,板蓝根样品中存在其他成分促进精氨酸、腺苷的P-gp外排作用。结论:板蓝根药材中精氨酸和腺苷能够被小肠细胞较好地吸收,存在吸收转运方向差异性,且都受到P-gp外排作用。

Absorption and Transport Characteristics Analysis of Effective Components from Isatidis Radix in Caco-2 Cell

Objective: To study absorption and transport characteristics of effective components from Isatidis Radix in Caco-2 cell. Method: Safety of drug candidates from Isatidis Radix was evaluated by establishing absorption and transport model of Caco-2 cells,which were qualified beforehand with three indexes including trans-epithelial electrical resistance(TEER),alkaline phosphatase activity and permeability of sodium fluorescein.Several influential factors for absorption and transport process of effective components from Isatidis Radix had been investigated,such as concentration,time,temperature,pH value and inhibitors.HPLC was set up for determination of these compounds,their apparent permeability coefficients(P_app) and efflux rates were calculated. Result: Arginine and adenosine from Isatidis Radix could be absorbed well by absorptive cells of small intestine with P_app of 1.01×10^-6-1.35×10^-6 cm·s^-1 and 0.40×10^-6-0.71×10^-6 cm·s^-1,respectively.Two kinds of ingredients show good permeability in Caco-2 cell,following by absorbed degree of arginine with rate of>50%,better than that of adenosine.Efflux rates(ER) of arginine and adenosine from Isatidis Radix extract were between 1.0-2.0,indicating P-glycoprotein(P-gp) efflux effect existed in absorption and transport process of these two compounds.And other compounds from Isatidis Radix could enhance efflux effect of P-gp on absorption of arginine and adenosine. Conclusion: Arginine and adenosine from Isatidis Radix can be absorbed well by absorptive cells of small intestine,and directional differences of absorption and transport process in Caco-2 cells are found out with P-gp efflux effect.