P-glycoprotein-mediated in vitro biliary excretion in sandwich-cultured rat hepatocytes.
P-glycoprotein-mediated in vitro biliary excretion in sandwich-cultured rat hepatocytes.
复制标题
DOI：
--
复制
发表时间：
2001-10
期刊：
影响因子：
--
通讯作者：
Pieter Annaert;Ryan Z. Turncliff;Catherine Booth;D. Thakker;Kim L. R. Brouwer
中科院分区：
文献类型：
--
作者：
Pieter Annaert;Ryan Z. Turncliff;Catherine Booth;D. Thakker;Kim L. R. Brouwer
最近，体外培养（SC）大鼠肝细胞已被用作体外模型，以评估药物和外源性物质的胆汁排泄。本研究的目的是验证SC大鼠肝细胞用于P-糖蛋白（P-gp）介导的胆汁药物排泄的体外评估。选择特异性和荧光P-gp底物罗丹明123（Rh 123）和P-gp底物地高辛作为模型化合物。在SC大鼠肝细胞中定量标准和无Ca（2+）条件下的Rh 123和地高辛蓄积以及Rh 123外排，以测定体外小管网络中的底物分泌。P-gp在这些化合物的胆汁排泄中的主要作用通过使用强效P-gp抑制剂GF 120918的抑制实验证实。肝细胞培养条件，包括培养基类型和培养时间，显着影响Rh 123胆汁排泄。P-gp表达，通过Western blot评估，随着培养时间的增加。细胞培养基中地塞米松（P-gp的体内诱导剂）浓度范围为0.01至1 μ M，不影响P-gp表达或Rh 123胆汁排泄。根据SC大鼠肝细胞数据预测的Rh 123和地高辛胆汁清除率值与体内和离体灌注大鼠肝脏研究中报告的值一致。总之，本研究的结果证明了SC大鼠肝细胞作为研究和预测P-gp底物胆汁排泄的体外模型的实用性。
Recently, sandwich-cultured (SC) rat hepatocytes have been used as an in vitro model to assess biliary excretion of drugs and xenobiotics. The purpose of the present study was to validate the use of SC rat hepatocytes for the in vitro assessment of P-glycoprotein (P-gp)-mediated biliary drug excretion. The specific and fluorescent P-gp substrate rhodamine 123 (Rh123) and the P-gp substrate digoxin were selected as model compounds. Rh123 and digoxin accumulation and Rh123 efflux under standard and Ca(2+)-free conditions were quantified in SC rat hepatocytes to determine substrate secretion into canalicular networks in vitro. The major role of P-gp in the biliary excretion of these compounds was confirmed by inhibition experiments with the potent P-gp inhibitor GF120918. Hepatocyte culture conditions, including media type and time in culture, significantly affected Rh123 biliary excretion. P-gp expression, as assessed by Western blot, was increased with culture time. Dexamethasone (an in vivo inducer of P-gp) concentrations ranging from 0.01 to 1 microM in the cell culture medium did not influence P-gp expression or Rh123 biliary excretion. Rh123 and digoxin biliary clearance values, predicted from SC rat hepatocyte data, were consistent with values reported in vivo and in isolated perfused rat liver studies. In conclusion, the results of this study demonstrate the utility of SC rat hepatocytes as an in vitro model to study and predict the biliary excretion of P-gp substrates.