Endo T, Kimura O, Sasaya M, Takada M, Sakata M
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Japan.
Biol Pharm Bull. 1995 Dec;18(12):1689-93. doi: 10.1248/bpb.18.1689.
Effects of sodium ions (Na+) and metabolic inhibitors on cadmium (Cd) uptake were investigated in LLC-PK1 cells derived from pig kidney under the nontoxic conditions of Cd. The inoculated cells became confluent on day 7 after the logarithmic growth phase. The initial uptake of Cd (1-60 microM) by the confluent cells (day 7) was assayed. The relationship between the Cd uptake at 37 degrees C and Cd concentration was nonlinear, but the relationship at 4 degrees C was linear. Subtraction of the Cd uptake at 4 degrees C from that at 37 degrees C showed a saturable uptake against the Cd concentration. By the Eadie-Hofstee analysis of saturable uptake, Km and Vmax were 24.6 microM and 164 pmol Cd/mg protein/min, respectively. The uptake of Cd by the cells was significantly decreased by ouabain and the metabolic inhibitors, and by the replacement of Na+ with potassium or choline ion in the incubation medium. These results suggest that Cd is incorporated into the confluent LLC-PK1 cells not only by simple diffusion but also by the carrier-mediated transport involved in Na(+)- and energy-dependent process(es).
在镉无毒的条件下,研究了钠离子(Na+)和代谢抑制剂对猪肾来源的LLC-PK1细胞摄取镉(Cd)的影响。接种的细胞在对数生长期后的第7天汇合。测定了汇合细胞(第7天)对镉(1-60 microM)的初始摄取。37℃下镉摄取与镉浓度之间的关系是非线性的,但4℃下的关系是线性的。从37℃下的镉摄取量中减去4℃下的镉摄取量,显示出对镉浓度的饱和摄取。通过对饱和摄取的伊迪-霍夫斯泰分析,Km和Vmax分别为24.6 microM和164 pmol Cd/mg蛋白质/分钟。哇巴因、代谢抑制剂以及在孵育培养基中用钾离子或胆碱离子替代钠离子,均可显著降低细胞对镉的摄取。这些结果表明,镉不仅通过简单扩散进入汇合的LLC-PK1细胞,还通过参与钠(+)和能量依赖过程的载体介导转运进入细胞。