鸡小肠原位单向灌流法和MDCK细胞系测定美托洛尔渗透性的影响因素  

Influence factors of metoprolol permeability determination in MDCK cell lines and chicken intestine

作　　者：郭荔[1] 李香秀 何方[1] 刘洋 张瑜娟 黄金虎[1] 王丽平[1] GUO Li;LI Xiangxiu;HE Fang;LIU Yang;ZHANG Yujuan;HUANG Jinhu;WANG Liping(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095

出　　处：《南京农业大学学报》2019年第3期505-512,共8页Journal of Nanjing Agricultural University

基　　金：国家重点研发计划项目(2016YFD0501309)

摘　　要：[目的]本试验旨在完善禽用药物生物药剂学分类系统(BCS)中药渗透性的准确测定方法。[方法]选取高渗透性内参药物美托洛尔,通过鸡小肠原位单向灌流法和体外过表达鸡P-gp的MDCK-chAbcb1单层细胞模型,探讨pH值(5、6和7)、药物浓度(4、40和400μg·mL^(-1))和小肠灌流部位(十二指肠、空肠和回肠)对美托洛尔渗透性测定的影响,为禽用药物BCS分类中渗透性测定方法的建立奠定基础。[结果]比较小肠不同部位在各自生理pH值条件下对美托洛尔的有效渗透系数(P_(eff))值时,发现回肠部位测得的P_(eff)最高(P<0.01),分别为十二指肠和空肠部位测得的2.2倍和2.3倍;原位灌流试验结果显示,灌流液pH值升高会增强药物在回肠的P_(eff)值,pH7时的P_(eff)值(1.35×10^(-4) cm·s^(-1))显著高于pH5时(0.72×10^(-4) cm·s^(-1)),同样在MDCK细胞中,随着pH值升高,药物渗透系数(P_(app))值均极显著升高(P<0.01);进一步采用受pH影响较小的空肠段进行灌流试验探讨不同药物浓度对渗透性测定的影响,发现增加肠灌流液中美托洛尔的浓度会极显著增加其P_(eff)值(P<0.01),4、40和400μg·mL^(-1)美托洛尔的P_(eff)分别为-0.48×10^(-4)、0.27×10^(-4)和3.04×10^(-4) cm·s^(-1),且在MDCK和MDCK-chAbcb1细胞测定的结果也显示,40和400μg·mL^(-1)美托洛尔的P_(app)值均显著高于4μg·mL^(-1)的(P<0.05),而40、400μg·mL^(-1)美托洛尔的P_(app)值间无显著差异(P>0.05)。[结论]机体内环境的pH值、药物浓度以及小肠灌流部位均可以影响体内外试验模型中药物渗透性测定的结果,故在建立药物渗透性测定方法时应充分考虑这些因素。[Objectives]The aim of the study is to develop the method to accurately determine the drug permeability for establishing Biopharmaceutics Classification System(BCS)of veterinary drugs.[Methods]In this study,metoprolol was selected as the high-permeability internal reference to study the effects of different intestinal segments(duodenum,jejunum and ileum),pH(5,6 and 7)and concentrations(4,40 and 400μg·mL^-1)of the metoprolol on the determination of drug permeability by in situ single-pass intestinal perfusion model in chickens and MDCK cell lines,respectively.[Results]The results from chicken in situ perfusion model showed that the ileum epithelial permeability(P eff)values of metoprolol in ileum were 2.2 and 2.3 times higher than those in duodenal and jejunum(P<0.01),respectively.The P eff of metoprolol at pH7 was significantly higher than that of metoprolol at pH5(0.72×10^-4 cm·s^-1 vs.1.35×10^-4 cm·s^-1).The same trend was found in MDCK cells that the permeability of metoprolol(P app)significantly increased under high pH value(P<0.01).Further we selected jejunum segment for the perfusion experiment of metoprolol at different concentrations.The results showed that the increase of perfusion drug concentration caused significantly higher P eff(P<0.01).The P eff values of metoprolol were-0.48×10-4,0.27×10^-4 and 3.04×10^-4 cm·s^-1 at different concentrations of 4,40 and 400μg·mL^-1,respectively.While in MDCK and MDCK-chAbcb1 cells,P app values of metoprolol at 4 and 400μg·mL^-1 were significantly higher than that of metoprolol at 4μg·mL^-1(P<0.05),however,there was not significant difference between P app value of metoprolol at 40 and 400μg·mL^-1(P>0.05).[Conclusions]In summary,the main factors involved in the drug permeability in chickens are related with the perfusion regions of the intestinal tract,local pH value and drug concentrations,which should be paid much attention to when determining the drug permeability by in vitro or in vivo method.

关 键 词：美托洛尔 鸡小肠原位单向灌流 MDCK细胞单层模型 渗透性 

分 类 号：S831.7[农业科学—畜牧学]