体外气血屏障通透性筛检模型的建立及应用测评  

Construction and evaluation of an in vitro alveolar-capillary barrier model for permeability screening

作　　者：赵康峰[1] 宋予晴 董力[1] 张兴 戴洪兴[3] 张宏伟[1] 鲁波[1] 白雪涛[1] ZHAO Kang-feng;SONG Yu-qing;DONG Li;ZHANG Xing;DAI Hong-xingZHANG Hong-wei;LU Bo;BAI Xue-tao(National Institute of Environmental Health,Chinese Center for Disease Control and Prevention,Beijing 100021,China;Peking Union Medical College,Beijing 100730,China;College of Environment and Energy Engineering,Beijing University of Technology,Beijing 100124,China)

机构地区：[1]中国疾病预防控制中心环境与健康相关产品安全所,北京100021 [2]北京协和医学院,北京100730 [3]北京工业大学环境与能源工程学院,北京100124

出　　处：《毒理学杂志》2019年第5期346-350,356,共6页Journal of Toxicology

基　　金：国家自然科学基金(81372949);中国疾病预防控制中心青年科研基金(2016A206)

摘　　要：目的建立气血屏障(ACB)通透性筛检细胞模型,并对其应用效能进行测评。方法以底膜孔径为1μm的细胞插板(12孔板匹配)为模型构建载体,采用肺毛细血管内皮细胞(PCECs)和A549细胞共培养方式建立ACB模型。测定跨内皮细胞电阻(TEER)评估模型的屏障特点。荧光黄(LY)作为指示剂,利用建立的模型进行纳米氧化铝(nano-Al2O3)(片状和短棍状)、纳米氧化锌(nano-ZnO)和纳米硫化铅(nano-PbS) 4种纳米材料和硫酸铝[Al2(SO4)3]、乙酸锌[Zn(CH3COO)2]、硝酸铅[Pb(NO3)2]3种化学物的通透性实验,计算通透系数(Pe)用来表示上述物质引起的ACB通透性改变情况。结果建立了由PCECs和A549细胞共同生长于细胞插板底膜(A549和PCECs分别生长于底膜内外表面)的ACB细胞通透性筛检模型,所建立模型的TEER峰值为(97. 2±0. 6)Ω·cm^2。实验测得片状nano-Al2O3、短棍状nano-Al2O3、nano-ZnO、nano-PbS、Al2(SO4)3、Zn(CH3COO)2和Pb(NO3)2对模型Pe分别是(2. 02±0. 31)×10^-3、(1. 49±0. 16)×10^-3、(1. 04±0. 08)×10^-3、(1. 03±0. 16)×10^-3、(1. 55±0. 21)×10^-3、(1. 01±0. 06)×10^-3和(1. 32±0. 06)×10^-3cm/min,其中片状nano-Al2O3、短棍状nano-Al2O3、Al2(SO4)3和Pb(NO3)24种物质Pe值明显高于模型对照组Pe值(1. 02±0. 03)×10^-3cm/min,差异有统计学意义(P<0. 05)。结论利用建立的ACB通透性筛检模型,可用于对潜在屏障通透性功能影响物质(纳米材料及一般化学物)进行快速筛查。Objective To construct an alveolar-capillary barrier(ACB) permeability screening model and to evaluate its effectiveness. Methods Pulmonary capillary endothelial cells(PCECs) and A549 cells(human alveolar adenocarcinoma cells) were co-cultured in transparent hanging cell inserts(PET membrane,pore size 1. 0 μm) matched with 12-well plates to build up in vitro ACB permeability screening model. Transendothelial electrical resistance(TEER) was measured to evaluate the barrier characteristics of the model. Then,the model was used to test the influence of sheet and short rod-shaped nano-aluminium oxide(nano-Al2 O3),nano-zinc oxide(nano-ZnO),nano-lead sulfide(nano-PbS),aluminium sulfate(Al2(SO4)3),zinc acetate(Zn(CH3 COO)2) and lead nitrate(Pb(NO3)2) on ACB permeability. Permeability coefficient(Pe) of LY(as tracer agent) was calculated to indicate the degree of permeability change. Result ACB permeability screening model through PCECs-A549 cells co-culturing in transparent hanging cell insert was built up,which A549 cells and PCECs grew on the internal and outside surface separately. The peak TEER value of the model was97. 2±0. 6 Ω·cm^2. The Pe values of sheet and short rod shaped nano-Al2 O3,nano-ZnO,nano-PbS,Al2(SO4)3,Zn(CH3 COO)2 and Pb(NO3)2 were calculated(2. 02±0. 31) ×10^-3,(1. 49±0. 16) ×10^-3,(1. 04±0. 08) ×10^-3,(1. 03±0. 16) ×10^-3,(1. 55±0. 21) ×10^-3,(1. 01±0. 06) ×10^-3 and(1. 32±0. 06) ×10^-3 cm/min. Among them Pe values of sheet and short rod shaped nano-Al2 O3,Al2(SO4)3 and Pb(NO3)2 were significantly higher than that of the control group [(1. 02±0. 03) ×10^-3 cm/min]. Conclusion In vitro ACB permeability screening model constructed is efficient for rapid screening for potential permeability-affected materials including nanomaterials and general chemicals.

关 键 词：气血屏障 体外模型 通透性改变 快速筛检 

分 类 号：R114[医药卫生—卫生毒理学]