Abstract：
An understanding of the quantitative relationship between bile canaliculus (BC) dynamics and the disruption of tight junctions (TJs) during drug-induced intrahepatic cholestasis may lead to new strategies aimed at drug development and toxicity testing. To investigate the relationship between BC dynamics and TJ disruption, we retrospectively analyzed the extent of TJ disruption in response to changes in the dynamics of BCs cultured with entacapone (ENT). Three hours after adding ENT, the ZO-1-negative BC surface area ratio became significantly higher (4.1-fold) than those of ZO-1-positive BCs. Based on these data, we calculated slopes of surface area changes, m, of each ZO-1-positive and ZO-1-negative BC. BCs with m ≤ 15 that fell within the 95% confidence interval of ZO-1-positive BCs were defined as ZO-1-positive. To validate this method, we compared the frequency of ZO-1-positive BCs, FZ , with that of BCs with m ≤ 15, FT , in culture using drugs that regulate TJ, or induce intrahepatic cholestasis. FT values were correlated with FZ under all culture conditions (R2 = .99). Our results indicate that the magnitude of BC surface area changes is a factor affecting TJ disruption, suggesting that maintaining TJ integrity by slowing BC dilation inhibits cell death.
摘要：
了解药物诱导的肝内胆汁淤积过程中胆管（BC）动力学与紧密连接（TJs）破坏之间的定量关系可能会导致针对药物开发和毒性测试的新策略。为了研究BC动力学与TJ破坏之间的关系，我们回顾性分析了与恩他卡朋(ENT)培养的BCs动力学变化有关的TJ破坏程度.添加ENT后三小时，ZO-1阴性BC表面积比ZO-1阳性BC的表面积明显高于（4.1倍）。基于这些数据,我们计算了表面积变化的斜率，m,每个ZO-1阳性和ZO-1阴性BC。m≤15且在ZO-1阳性BCs的95%置信区间内的BCs被定义为ZO-1阳性。要验证此方法，我们比较了ZO-1阳性BCs的频率，FZ,对于m≤15的BC，FT，在使用调节TJ的药物的培养中，或诱发肝内胆汁淤积。在所有培养条件下,FT值与FZ相关(R2=.99)。我们的结果表明，BC表面积变化的大小是影响TJ破坏的一个因素，表明通过减缓BC扩张来维持TJ完整性抑制细胞死亡。
