Malik J K, Schwarz L R, Wiebel F J
Chem Biol Interact. 1983 Jul 1;45(1):29-42. doi: 10.1016/0009-2797(83)90040-6.
The present studies were aimed at evaluating procedures for assessing the effect of chemicals on the integrity of the plasma membrane in continuous cell cultures. The degree of membrane damage was monitored by determining the 'leakage' of alpha-[3H]aminoisobutyric acid ([3H]AIB) and [14C]deoxy-2-fluoro-D-glucose ([14C]FdG) from the prelabelled cells. These parameters were compared to the loss of lactate dehydrogenase (LDH) from the cells and the decrease in the intracellular level of K+. Triton X-100, sodium dodecylsulfate (SDS), phospholipase C and nystatin which are known to affect membranes by different mechanisms served as test agents. In parallel, we monitored the effects of the chemicals on the viability of the cells. The following results were obtained: (1) The two radioactive markers [3H]AIB and [14C]FdG were found to be suitable to probe for damages of the plasma membrane in a variety of continuous cell lines which differ widely in their phenotype, rate of growth and degree of differentiation. (2) The leakage of the two markers could conveniently be monitored by double labelling techniques. (3) The loss from the cells of the 3 markers of smaller molecular size, K+, [3H]AIB, [14C]FdG, differed considerably depending on the test agent used. (4) Intracellular K+ level and [3H]AIB leakage generally appeared to follow a similar pattern, whereas [14C]FdG leakage may have shown a distinctly different response. (5) The leakage of LDH was an insensitive indicator for membrane damage. (6) No clear relationship was detectable between a particular leakage pattern of the markers and the loss of cellular viability.
本研究旨在评估在连续细胞培养中评估化学物质对质膜完整性影响的程序。通过测定预标记细胞中α-[³H]氨基异丁酸（[³H]AIB）和[¹⁴C]脱氧-2-氟-D-葡萄糖（[¹⁴C]FdG）的“泄漏”来监测膜损伤程度。将这些参数与细胞中乳酸脱氢酶（LDH）的损失以及细胞内K⁺水平的降低进行比较。已知通过不同机制影响膜的 Triton X-100、十二烷基硫酸钠（SDS）、磷脂酶C和制霉菌素用作测试剂。同时，我们监测了化学物质对细胞活力的影响。获得了以下结果：（1）发现两种放射性标记物[³H]AIB和[¹⁴C]FdG适用于探测多种连续细胞系中的质膜损伤，这些细胞系在表型、生长速率和分化程度上有很大差异。（2）两种标记物的泄漏可以通过双标记技术方便地进行监测。（3）细胞中较小分子大小的三种标记物K⁺、[³H]AIB、[¹⁴C]FdG的损失因所用测试剂的不同而有很大差异。（4）细胞内K⁺水平和[³H]AIB泄漏通常似乎遵循相似的模式，而[¹⁴C]FdG泄漏可能表现出明显不同的反应。（5）LDH的泄漏是膜损伤的不敏感指标。（6）在标记物的特定泄漏模式与细胞活力丧失之间未发现明显关系。
