大鼠原位肠—肝血管灌流模型在药物吸收方面的优化与验证

发布时间：2018-05-23 11:21

  本文选题：大鼠原位肠-肝血管灌流模型 + Caco-2细胞模型　； 参考：《广东药学院》2012年硕士论文

【摘要】：目的：优化大鼠原位肠-肝血管灌流模型，并验证该模型在预测药物吸收方面的可行性。 方法：对灌流条件进行优化，分别对Krebs-Ringer缓冲液（K-R液）加牛血清白蛋白和甘露醇、K-R液加牛血清白蛋白和右旋糖酐T-40二者进行对比，找出最佳的灌流介质，然后对灌流液中牛血清白蛋白的含量进行优化；建立三种生物碱（氧化苦参碱、苦参碱和槐果碱）和六种有机酸（绿原酸、咖啡酸、阿魏酸、肉桂酸、甘草酸和甘草次酸）共九种化合物在灌流液、大鼠血浆和HBSS中的LC-MS/MS分析方法；以优化的大鼠原位肠-肝血管灌流模型研究九种化合物的灌流吸收率；通过整体动物实验研究九种化合物在大鼠体内的药动学特征和绝对生物利用度（Fabs）；建立Caco-2细胞模型研究九种化合物的双向转运过程并计算表观渗透系数（Papp）；运用灰色关联分析法对九种化合物的灌流吸收率、Fabs和正向表观渗透系数（PappA→B）进行相关性分析。 结果：优化的大鼠原位肠-肝血管灌流模型能更经济、准确地预测药物的吸收情况；氧化苦参碱、苦参碱、槐果碱、绿原酸、咖啡酸、阿魏酸、肉桂酸、甘草酸和甘草次酸在优化的灌流模型中的吸收率分别为8.30%、14.03%、12.03%、1.37%、2.68%、59.00%、85.59%、0.47%和88.73%；在大鼠体内的Fabs分别为15.69%、53.40%、39.63%、3.59%、14.70%、64.91%、81.15%、3.04%和96.72%；在Caco-2细胞模型中的PappA→B分别为（6.39±0.74）×10-6cm/s、（10.7±0.17）×10-6cm/s、（7.35±0.76）×10-6cm/s、（0.66±0.04）×10-6cm/s、（5.05±0.66）×10-6cm/s、（10.24±1.58）×10-6cm/s、（24.30±3.82）×10-6cm/s、（0.28±0.06）×10-6cm/s和（30.57±4.69）×10-6cm/s；灰色关联分析表明灌流吸收率和Fabs的关联度为0.7194，，PappA→B与Fabs的关联度为0.7515，二者没有显著性差异（P＞0.05）。 结论：优化的大鼠原位肠-肝血管灌流模型在药物吸收研究领域具有良好的应用前景。
[Abstract]:Aim: to optimize the in situ intestinal-hepatic vascular perfusion model in rats and to verify the feasibility of the model in predicting drug absorption. Methods: the perfusion conditions were optimized and the optimal perfusion medium was found by comparing the Krebs-Ringer buffer solution with bovine serum albumin and mannitol K-R solution with bovine serum albumin and dextran T-40. Three alkaloids (oxymatrine, matrine and sophorine) and six organic acids (Lv Yuan acid, caffeic acid, ferulic acid, cinnamic acid) were established. The method of LC-MS/MS analysis of nine compounds in perfusate, rat plasma and HBSS was used to study the perfusion absorption rate of nine compounds in the model of in situ enteric-hepatic vascular perfusion in rats with glycyrrhizic acid and glycyrrhetinic acid (glycyrrhizic acid). The pharmacokinetic characteristics and absolute bioavailability of nine compounds in rats were studied by whole animal experiment, the bidirectional transport process of nine compounds was studied by Caco-2 cell model and the apparent osmotic coefficient was calculated. Correlation analysis was carried out to analyze the correlation between the perfusion absorptivity (Fabs) and the forward apparent osmotic coefficient (PappA B) of nine compounds. Results: the optimized in situ enterohepatic vascular perfusion model could be more economical and accurate in predicting drug absorption, oxymatrine, sophoricine, Lv Yuan acid, caffeic acid, ferulic acid, cinnamic acid, 鐢樿崏閰稿拰鐢樿崏娆￠吀鍦ㄤ紭鍖栫殑鐏屾祦妯″瀷涓�鐨勫惛鏀剁巼鍒嗗埆涓�8.30%,14.03%,12.03%,1.37%,2.68%,59.00%,85.59%,0.47%鍜

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