体外模拟单室血管外给药染毒模型的构建及其与经典体外染毒模型在硝酸镧诱导HepG2细胞死亡中的比较  

Construction of an in vitro simulated one compartment extravascular administration model and comparisons with a classic in vitro administration model in lanthanum nitrate induced HepG2 cell death

作　　者：付大维 付于津 闫赖赖[2] 陈洁 刘芷毓 付娟玲[1] 姚碧云[1] 郝卫东[1] 赵鹏[1] FU Dawei;FU Yujin;YAN Lailai;CHEN Jie;LIU Zhiyu;FU Juanling;YAO Biyun;HAO Weidong;ZHAO Peng(Department of Toxicology,Beijing Key Laboratory of Toxicology Research and Risk Assessment for Food Safety,Peking University,Beijing 100191,China;Department of Laboratorial Science and Technology in Preventive Medicine,School of Public Health,Peking University,Beijing 100191,China)

机构地区：[1]北京大学公共卫生学院毒理学系,食品安全毒理学研究与评价北京市重点实验室,北京100191 [2]北京大学公共卫生学院卫生检验学系,北京100191

出　　处：《中国药理学与毒理学杂志》2025年第4期285-295,I0001,I0002,共13页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81370079),国家自然科学基金(81001253);北京市自然科学基金(7132122)。

摘　　要：目的以硝酸镧﹝La(NO_(3))_(3)﹞为受试物,构建体外模拟单室血管外给药染毒模型,并探究其与经典体外染毒模型在La(NO_(3))_(3)诱导Hep G2细胞死亡中的差异。方法根据毒动学房室模型理论,自主设计由储液室、混合室、染毒室和废液室组成的染毒装置,采用蠕动泵软管连接各室以单向和定速传输液体,根据预设的吸收半衰期(T_(1/2a))和消除半衰期(T_(1/2))等毒动学参数构建La(NO_(3))_(3)体外模拟单室血管外给药染毒模型。采用电感耦合等离子体质谱仪测定不同时间点染毒室中La(NO_(3))_(3)含量。采用PKsolver和Graph Pad Prism 8.0软件对La(NO_(3))_(3)浓度-时间曲线进行分析。通过比较毒动学参数实测值和根据假定的T_(1/2a)和T_(1/2)计算得到的理论值对染毒模型进行评估。以模拟单室血管外给药染毒模型和经典体外染毒模型对Hep G2细胞给予La(NO_(3))_(3)染毒处理,采用Hoechst 33342/碘化丙啶染色测定细胞死亡率。结果在峰值浓度(Cmax)3.91~1000.00μmol·L^(-1)范围内,体外模拟单室血管外染毒模型实测的La(NO_(3))_(3)浓度-时间曲线几乎与相应理论曲线一致,测量值与理论值呈良好线性相关性(r均>0.9980);包括消除速率常数(Ke)、T1/2、吸收速率常数(Ka)、T_(1/2a)、达峰时间(T_(max))、C_(max)、清除率(CL)和曲线下面积(AUC_(0-∞))在内的各毒动学参数实测值均与相应理论值接近;各实验组浓度-时间曲线拟合系数R2均>0.9900,符合单室血管外给药的房室模型。模拟单室血管外给药染毒模型中,La(NO_(3))_(3)各剂量组均未观察到Hep G2细胞出现明显死亡。经典体外染毒模型中,La(NO_(3))_(3)0.500 mmol·L^(-1)组细胞死亡率显著高于溶剂对照组;0.119和0.243 mmol·L^(-1)组未见明显细胞死亡。Cmax或C染毒为0.500 mmol·L^(-1)时,经典体外染毒诱导Hep G2细胞死亡率显著高于模拟单室血管外给药染毒。AUC相等时,2种染毒模型间La(NO_(3))_(3)诱导HeOBJECTIVE To establish an in vitro simulated one compartment extravascular adminis⁃tration model with lanthanum nitrate as the test substance,and explore the differences between this model and the classic in vitro administration model in lanthanum nitrate induced HepG2 cell death.METHODS An in vitro administration device was designed based on compartment model theories which consisted of four functional chambers:the liquid storage chamber,mixing chamber,toxicant exposure chamber,and waste liquid receiving chamber.The four chambers were connected by peristaltic pump hoses.The peristaltic pumps were employed to ensure unidirectional and constant speed trans⁃mission of liquid between these chambers.According to the preset toxicokinetic parameters such as T_(1/2a)and T_(1/2),an in vitro simulated one compartment extravascular administration model of lanthanum nitrate was constructed using the device.The content of lanthanum nitrate in the toxicant exposure chamber at different time points was measured using inductively coupled plasma mass spectrometry.The concentration-time curves of lanthanum nitrate were analyzed using PKsolver and GraphPad Prism 8.0 software.The constructed in vitro simulated one compartment extravascular administration model was evaluated by comparing the measured and theoretical values of toxicokinetic parameters.HepG2 cells were treated with lanthanum nitrate in the in vitro simulated one compartment extravascular administration model and classic in vitro administration model,respectively,and cell death was measured using the Hoechst 33342/propidium iodide staining method.RESULTS Within the Cmax range of 3.91-1000.00μmol·L^(-1),the measured concentration-time curves of lanthanum nitrate in the toxicant expo⁃sure chamber almost conformed with the corresponding calculated theoretical curves(the correlation coefficients were all>0.9980).The measured values of toxicokinetic parameters,including Ke,T_(1/2),Ka,T_(1/2a),T_(max),C_(max),CL and AUC_(0-∞),were close to the corresponding theoret

关 键 词：体外模拟单室血管外给药染毒模型 经典体外染毒模型 毒动学 曲线下面积 硝酸镧 细胞死亡 

分 类 号：R99[医药卫生—毒理学]