白杨素及其衍生物在Caco-2细胞模型中的转运规律

Transport of Chrysin and Its Derivative across Caco-2 Monolayer Model

目的研究白杨素(chrysin,ChR)及其衍生物6,8-二-三氟甲基-7-乙酰氧基白杨素(dFMAChR)在人源结肠腺癌细胞系Caco-2细胞单层模型的转运规律。方法建立精确灵敏、重复性好的ChR及dFMAChR的高效液相色谱(HPLC)检测方法;MTT(四甲基偶氮唑盐)法测定ChR和dFMAChR对Caco-2细胞的毒性作用。体外培养Caco-2细胞,用Transwell建立单层细胞模型;确定ChR和dFMAChR转运的最佳pH条件。评价时间、浓度对dFMAChR和ChR的双向转运的影响,计算转运速率及表观渗透系数(Papp),并与阳性对照药普萘洛尔进行比较。结果 ChR和dFMAChR的转运最佳介质pH分别为6.0和6.5,两者的转运无论是Apical侧(A侧)到Basolateral侧(B侧)还是B侧到A侧,均具有时间依赖性和浓度依赖性。ChR和dFMAChR的平均Papp(A-B)分别是(1.60±0.15)×10-6 cm/s和(10.63±0.35)×10-6 cm/s,平均Papp(B-A)分别是(1.22±0.17)×10-6 cm/s和(10.43±0.28)×10-6 cm/s,两者Papp(A-B)>Papp(B-A),且Papp(A-B)/Papp(B-A)<2。结论在Caco-2模型中,dFMAChR和ChR均由被动扩散方式转运,dFMAChR的转运速率明显高于ChR,吸收接近普萘洛尔,属于吸收良好的化合物。

Objective To study the transport character of chrysin （ChR） and 6,8-ditrifluoromethyl-7-acetoxychrysin （dFMAChR） in Caco-2 monolayer model.Methods Precise,sensitive determination methods of ChR and dFMAChR by HPLC with good reproducibility were eatablished.MTT assay was used to determine the cell toxicity of ChR and dFMAChR to Caco-2 cells.Caco-2 cells were cultured in vitro and Caco-2 monolayer model was established in Transwell system.The optimal transportation mediator pH of dFMAChR and ChR were evaluated.The relationship of time and concentration with transportation of dFMAChR and ChR was evaluated by calculating transportation rate and apparent permeability coefficient （Papp） with propranolol as control.Results The optimal transportation mediator pH of dFMAChR and ChR were 6.5 and 6.0,respectively.The transport of both the A side to B side and B side to A side,has a time-and concentration-dependent manner.The average Papp （A-B） of ChR and dFMAChR were （1.60 ± 0.15）× 10-6 cm/s and （10.63±0.35） × 10-6 cm/s,the average Papp （B-A） of ChR and dFMAChR were （1.22±0.17） × 10-6 cm/s and （10.43±0.28） × 10-6 cm/s.Both ChR and dFMAChR had Papp （A-B） ＞ Papp （B-A） and Papp （A-B） /Papp （B-A） ＜ 2.Conclusion In the Caco-2 model,both dFMAChR and ChR obey passive diffusion transport.The transportation rate of dFMAChR is higher than ChR.The absorption of dFMAChR is approach of propranolol,and dFMAChR belongs to a compound with good absorption.