Chieli E, Santoni Rugiu E, Cervelli F, Paolicchi A, Tongiani R
Dipartimento de Biomedicina Sperimentale, Università di Pisa, Italy.
Cell Biol Toxicol. 1993 Jul-Sep;9(3):235-41. doi: 10.1007/BF00755602.
To test the activity of P-170 glycoprotein in isolated hepatocytes, a method has been developed employing the fluorescent dye rhodamine 123 (R-123). The uptake of R-123 by both freshly isolated and 4-hr-plated hepatocytes depends on dye concentration, time of incubation, and cell number. The efflux of R-123 from cells is inhibited by sodium azide and by verapamil. In standard conditions the efficiency of efflux of R-123 from cells correlates with the relative amount of immunoblottable glycoprotein. The method has been applied to detection of P-170 activity in hepatocytes from animals of different ages as well as from carcinogen-treated animals. The proposed assay appears a simple and adequate tool for the functional assessment of multidrug transporter in liver.
为检测分离的肝细胞中P - 170糖蛋白的活性，已开发出一种使用荧光染料罗丹明123（R - 123）的方法。新鲜分离的肝细胞和培养4小时的肝细胞对R - 123的摄取取决于染料浓度、孵育时间和细胞数量。叠氮化钠和维拉帕米可抑制R - 123从细胞中的外排。在标准条件下，R - 123从细胞中外排的效率与免疫印迹可检测的糖蛋白的相对量相关。该方法已应用于检测不同年龄动物以及经致癌物处理动物的肝细胞中P - 170的活性。所提出的检测方法似乎是一种用于肝脏中多药转运蛋白功能评估的简单且合适的工具。
