田蓟苷在Caco-2细胞模型中的吸收特性研究

Intestional absorption and mechanism of tiliani in Caco-2 cell model

目的:研究田蓟苷在Caco-2细胞模型的吸收特性。方法:体外培养的人小肠上皮细胞模型Caco-2,高效液相色谱法测定田蓟苷含量,进行田蓟苷在小肠上皮细胞的摄取、跨膜转运及外排研究。探讨时间、pH、药物浓度及抑制剂维拉帕米、叠氮化钠、2,4-二硝基酚、根皮苷和乳糖对Caco-2细胞摄取田蓟苷的影响。结果:田蓟苷在Caco-2细胞中的摄取呈时间依赖性;在4～16 mg.L-1,摄取呈浓度依赖性,符合被动扩散过程;在pH 5.0～8.0,酸性条件下有利于药物吸收;加入维拉帕米后,田蓟苷的细胞摄取量为(1.545±0.010)mg.g-1,与对照组相比,显著增高(P<0.05);加入叠氮化钠、2,4-二硝基酚以及根皮苷后,田蓟苷的细胞摄取量分别为(0.994±0.003),(1.174±0.030),(1.098±0.021)mg.g-1,与对照组相比,显著降低(P<0.05);加入乳糖后,田蓟苷的细胞摄取量为(1.470±0.025)mg.g-1,与对照组相比,差异无显著性;药物从basolateral(B,基底面)到apical(A,肠腔面)的渗透系数Papp大于A到B(1.10倍)。结论:P-gp以及Na+依赖葡萄糖转运载体1(SGLT1)参与田蓟苷的转运过程;田蓟苷的吸收与乳糖酶根皮苷水解酶(LPH)无关;被动扩散和载体媒介转运共同参与田蓟苷细胞吸收过程。

Objective： To observe the uptake of tiliani in Caco-2 Cell.Method： A human intestinal epithelial cell model Caco-2 cell in vitro cultured was applied to study the kinetics of uptake,transport and efflux kinetics of tiliani at small intestine.The effecte of time,pH,drug concentration and inhibitors on the uptake of tiliani were investigated.The determination of tiliani was performed by HPLC.Result： Tiliani in Caco-2 cell uptake was time-dependent.tiliani in Caco-2 cell uptake was concentration-dependent at 4-16 mg·L^-1 consistent with passive diffusion process.The acid condition was good for the uptake of tiliani at pH 5-8.Compared with the control group,tiliani cell uptake was significantly higher after additional treatmeant with verapamil（1.545±0.010） mg·g^-1,（P〈0.05）,and tiliani cell uptake was significantly lower after additional treatmeanet with sodium azide（0.994±0.003） mg·g^-1（P〈0.05）,with 2,4-dinitrophenol（1.174±0.030） mg·g^-1（P〈0.05）,and with phloridzin（1.098±0.021） mg·g^-1（P〈0.05）.Compared with the control group,tiliani cell uptake was not significantly after additional treatmeant with lactose（1.470±0.025） mg·g^-1,Papp of Basolateral to Apical was much more than that of Apical to Basolateral（1.10 Fold）.Concluson： P-glycopro-teins and SGLT1 participate in the conveying process of tiliani in Caco-2 cells.The uptake of tiliani has no relationship to LPH.passive transport and carrier-mediated transport participate in the uptake process of tiliani in Caco-2 cells.