1.College of Traditional Chinese Medicine, Xinjiang Medical University;2.Xinjiang Uygur Autonomous Region Institute of Pharmaceutical Inspection
The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan)
摘 要：目的 本研究建立 Caco-2 单层细胞模型研究野鸦椿酸摄取和转运特性。方法 采用超高效液相色谱-三重四极杆质谱联用仪（UPLC- TQ-MS）测定野鸦椿酸的含量，考察不同时间、温度对其摄取的影响，在此基础上考察不同浓度、P-gp抑制剂、螯合剂和pH值对其双向转运的影响。结果 在37℃条件下，野鸦椿酸在Caco-2 细胞模型上180min时的摄取量为（8.38 ?.87）μg穖g-1。野鸦椿酸低、中、高浓度的表观渗透系数（Papp值）与浓度呈正相关，分别为（61.41 ?.92）?0-4、（146.90?4.91）?0-4、（167.18?.72）?0-4 cm穝-1，P-gp抑制剂和螯合剂对其Papp值无影响，弱酸性环境（pH6.00）可明显提高其Papp值，其外排率（ER）介于0.8~1.4之间。结论 在Caco-2细胞模型中野鸦椿酸跨膜渗透性良好，其主要吸收方式为被动扩散，不是P-gp的底物，且不存在细胞旁路转运。本研究可为含有野鸦椿酸的药物的体内肠道吸收提供实验依据。
ABSTRACT: Objective Caco-2 cell model was established to explore euscaphic acid uptake and transport mechanism in this study. Methods Ultra-high performance liquid chromatography-triple quadrupole-mass spectrometry(UPLC-TQ-MS) was employed to determine the content of euscaphic acid. The effects of different time and temperature on its uptake were investigated. Based on the results of the uptake studies, the effects of different concentrations, P-gp inhibitors, chelating agents and pH values on its bidirectional transport were explored. Results The uptake of euscaphic acid was (8.38 ?0.87) μg穖g-1 for 180 min on the Caco-2 cell model at 37 癈. The apparent permeability coefficient (Papp values) of euscaphic acid for low, medium and high concentrations were(61.41 ?.92)?0-4、(146.90?4.91)?0-4 and (167.18?.72)?0-4 cm穝-1 respectively, which were positively correlated with the concentrations. P-gp inhibitors and chelating agents had no effect on its Papp values. Weakly acidic environment (pH6.00) significantly increased its Papp value, and the efflux rate (ER) ranged from 0.8–1.4. Conclusion The above results indicate that euscaphic acid has good transmembrane permeability in Caco-2 cell model, and the uptake mode is mainly passive diffusion. It is not a substrate of P-gp and there is no cellular bypass transport. This research may provide an experimental basis for the in vivo intestinal absorption of medicines containing euscaphic acid.