巴戟天中有效成分在Caco-2细胞模型中的吸收转运

Absorption and Transport Characteristics of Effective Components of Morinda officinalis in Caco-2 Cell Model

目的:研究巴戟天中的有效成分水晶兰苷、去乙酰车叶草苷酸和耐斯糖在人源结肠癌细胞Caco-2单层模型中的吸收特性。方法:通过细胞培养技术建立Caco-2细胞单层模型,以细胞形态学特征、细胞跨膜电阻等指标验证和筛选模型。考察不同pH、不同质量浓度的水晶兰苷、去乙酰车叶草苷酸和耐斯糖对Caco-2细胞活力的影响,以确定给药pH和质量浓度。通过Caco-2细胞转运实验探究药物质量浓度、作用时间与P-糖蛋白抑制剂维拉帕米对水晶兰苷、去乙酰车叶草苷酸和耐斯糖吸收转运的影响。采用高效液相色谱法(HPLC)测定水晶兰苷、去乙酰车叶草苷酸和耐斯糖的累积转运量。结果:水晶兰苷、去乙酰车叶草苷酸和耐斯糖在Caco-2细胞单层模型中吸收良好,三者低质量浓度的双侧表观渗透系数(P_(app))明显大于中、高质量浓度;加入盐酸维拉帕米后,三者的P_(app)变化不大,说明这3个成分的小肠吸收不需要载体。去乙酰车叶草苷酸的液相色谱图中存在少量水晶兰苷,说明去乙酰车叶草苷酸存在首过效应。结论:巴戟天中水晶兰苷、去乙酰车叶草苷酸和耐斯糖3个特征性成分在跨膜转运过程中均以被动转运为主,且3个成分都存在自身质量浓度抑制现象。

Objective:To investigate the absorption characteristics of monotropein,desacetyl asperulosidic acid,and nystose of Morinda officinalis How.in the monolayer of human colon cancer cell line Caco-2 cells.Methods:The Caco-2 cell monolayer models were prepared by cell culture technique,verified,and screened out with cell morphological characteristics,transepithelial electrical resistance,and other indicators.The effects of pH values and concentrations of monotropein,desacetyl asperulosidic acid,and nystose on Caco-2 cell viability were examined to determine the proper pH and concentrations of dosage in the experiment.The effects of drug concentration,action time,and P-glycoprotein inhibitor verapamil on the absorption and transport of monotropein,desacetyl asperulosidic acid,and nystose were investigated by the Caco-2 cell transport experiment.The cumulative transport of monotropein,desacetyl asperulosidic acid,and nystose was determined by high-performance liquid chromatography(HPLC).Results:Monotropein,desacetyl asperulosidic acid,and nystose were well absorbed in the Caco-2 cell monolayer model,and the bilateral apparent permeability coefficients(P_(app))at low concentrations were significantly greater than those at medium and high concentrations.After the addition of verapamil hydrochloride,the P_(app) of the three components changed slightly,indicating that the carrier was not required in the intestinal absorption of these three components.A small amount of monotropein presented in the liquid chromatogram of desacetyl asperulosidic acid,indicating that desacetyl asperulosidic acid showed the first-pass effect.Conclusion:The three characteristic components of M.officinalis,monotropein,desacetyl asperulosidic acid,and nystose were mainly passively transported in the transmembrane transport,and all the three components showed inhibition of their own concentrations.