Shen Jie, Cross Stacy T, Tang-Liu Diane D S, Welty Devin F
Department of Pharmacokinetics and Drug Metabolism, Allergan, Inc., Irvine, California 92612, USA.
Pharm Res. 2003 Sep;20(9):1357-63. doi: 10.1023/a:1025789606885.
To evaluate the growth and barrier properties of an immortalized rat retinal endothelial cell line (TR-iBRB) maintained on permeable membrane for drug transport studies.
TR-iBRB cells were grown on permeable membrane filters. The effect of coating material on cell growth was investigated. Transport of [14C]-3-O-methyl-D-glucose (3-OMG), AGN 194716, AGN 195127, AGN 197075, acebutolol, alprenolol, atenolol, brimonidine, carbamazepine epoxide (CBZ-E), metoprolol, nadolol, rhodamine 123, and sotalol was measured across the cultured cell layer to determine the apparent permeability coefficients (Papp). Rhodamine 123 uptake into these cells in the presence of these test compounds was evaluated. Western blot was performed to detect the efflux transporter P-glycoprotein (P-gp). Bidirectional transport in MDR1-MDCK cell monolayers overexpressing the human P-gp was measured for AGN 197075.
TR-iBRB cells form confluent cell layers when grown on fibronectin-coated membrane and exhibit characteristic spindle-shaped morphology. A good correlation between Papp and cLogD (pH 7.4) of the compounds tested was observed, except for 3-OMG, AGN 197075, and rhodamine 123, which are substrates of carrier-mediated transport systems such as P-gp and a glucose transporter (GLUT1). When grown on permeable membrane, TR-iBRB cells expressed functional P-gp and GLUT1.
TR-iBRB cells, when grown on permeable membrane, provide a useful tool for predicting permeability across the BRB. The usefulness of this model for high-throughput screening and rank ordering of drug candidates intended for the back of the eye in treatment of ocular diseases needs further characterization upon correlation with in vivo data.
评估在可渗透膜上培养的永生化大鼠视网膜内皮细胞系（TR-iBRB）的生长及屏障特性，用于药物转运研究。
将TR-iBRB细胞接种于可渗透膜滤器上。研究包被材料对细胞生长的影响。测定[14C]-3-O-甲基-D-葡萄糖（3-OMG）、AGN 194716、AGN 195127、AGN 197075、醋丁洛尔、阿普洛尔、阿替洛尔、溴莫尼定、卡马西平环氧化物（CBZ-E）、美托洛尔、纳多洛尔、罗丹明123和索他洛尔透过培养细胞层的转运情况，以确定表观渗透系数（Papp）。评估在这些测试化合物存在下罗丹明123对这些细胞的摄取情况。进行蛋白质免疫印迹法检测外排转运体P-糖蛋白（P-gp）。测定AGN 197075在过表达人P-gp的MDR1-MDCK细胞单层中的双向转运。
TR-iBRB细胞在纤连蛋白包被的膜上生长时形成汇合细胞层，并呈现特征性的纺锤形形态。观察到所测试化合物的Papp与cLogD（pH 7.4）之间具有良好的相关性，但3-OMG、AGN 197075和罗丹明123除外，它们是载体介导转运系统如P-gp和葡萄糖转运体（GLUT1）的底物。当在可渗透膜上生长时，TR-iBRB细胞表达功能性P-gp和GLUT1。
TR-iBRB细胞在可渗透膜上生长时，为预测药物透过血视网膜屏障的通透性提供了有用的工具。该模型用于眼部疾病治疗中眼后段候选药物的高通量筛选和排序的实用性，在与体内数据相关联后还需要进一步表征。
