Akahori A, Jóźwiak Z, Gabryelak T, Gondko R
Department of Thermobiology, Institute of Biophysics, University of Lódź, Poland.
Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1999 Jul;123(3):209-15. doi: 10.1016/s0742-8413(99)00031-6.
The effect of zinc exposure on some properties of the carp erythrocyte membrane was studied in vitro. Red blood cells plasma membranes were separated from other cellular membranes using a combination of differential and density gradient centrifugation. The purity of obtained plasma membrane preparations was determined by measuring the activity of the marker enzymes. Electrophoretic patterns of the main erythrocyte membrane proteins excluded their degradation during the isolation and purification procedure. Carp erythrocyte membranes, obtained from cells previously incubated with increasing ZnSO4 concentrations, were used to elucidate the effect of zinc ions on their physical and biochemical properties. Using fluorescent probes: 12-AS and TMA-DPH, we found that zinc ions reduced the fluidity of the lipid bilayer, both in the middle and near the aqueous interface. Moreover, it was observed that zinc had no significant influence neither on the Na,K-ATPase activity nor on the thiol groups content in the erythrocyte membrane. We also detected that incubation of erythrocytes with zinc lead to the marked decrease of hemolytic resistance of the cells. Our studies demonstrate that zinc at higher concentrations may be toxic to carp erythrocytes causing changes in the membrane fluidity and hemolytic resistance.
在体外研究了锌暴露对鲤鱼红细胞膜某些特性的影响。使用差速离心和密度梯度离心相结合的方法从其他细胞膜中分离出红细胞质膜。通过测量标记酶的活性来确定所得质膜制剂的纯度。主要红细胞膜蛋白的电泳图谱排除了其在分离和纯化过程中的降解。从先前用浓度递增的硫酸锌孵育的细胞中获得的鲤鱼红细胞膜用于阐明锌离子对其物理和生化特性的影响。使用荧光探针12-AS和TMA-DPH,我们发现锌离子降低了脂质双层在中间和靠近水界面处的流动性。此外,观察到锌对红细胞膜中的钠钾ATP酶活性和硫醇基团含量均无显著影响。我们还检测到用锌孵育红细胞会导致细胞溶血抗性显著降低。我们的研究表明,较高浓度的锌可能对鲤鱼红细胞有毒,导致膜流动性和溶血抗性发生变化。