Reader S C, Foster P M
ICI PLC Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire, United Kingdom.
Toxicol Appl Pharmacol. 1990 Nov;106(2):287-94. doi: 10.1016/0041-008x(90)90248-s.
The present study was undertaken to evaluate the in vitro effects of four isomers of a known testicular toxicant, dinitroluene (DNT). Rat Sertoli or Sertoli-germ cell cocultures were treated, after 3 days in culture, with DNT isomers (0.01 to 100 microM) or 1,3-dinitrobenzene (1,3-DNB) for 24 hr. Cellular morphology, germ cell detachment (GCD) and lactate pyruvate production were used as sensitive effect markers of in vitro toxicity. Morphologically the Sertoli cell monolayer remained intact 24 hr after exposure to DMSO, 1,3-DNB, or DNT isomers. Some apparent cytotoxicity was observed at 100 microM 3,4-DNT: the monolayer was disrupted with extensive vacuolation of the Sertoli cells. Cocultures treated with concentrations of 50 microM DNT isomers closely resembled cells treated with 100 microM 1,3-DNB. GCD increased in a dose-dependent manner (0.01 and 10 microM DNT isomers) increasing between 2- and 10-fold over control. Both lactate and pyruvate production increased with rising concentrations of DNT isomers. The most sensitive effect was seen with 3,4-DNT (10 to 25 microM). In the case of 2,6-DNT, despite increases in GCD and lactate production, only a minimal increase in pyruvate was demonstrated. Overall, the ratio of lactate to pyruvate production declined with increasing doses of DNT. These results indicate that the four isomers of DNT directly affected Sertoli cell morphology and function, effects comparable to those seen with the Sertoli cell toxicant 1,3-DNB. Further, the data support the hypothesis that DNT may be a Sertoli cell toxicant.
本研究旨在评估一种已知的睾丸毒物二硝基甲苯（DNT）的四种异构体的体外效应。培养3天后，用DNT异构体（0.01至100微摩尔）或1,3 - 二硝基苯（1,3 - DNB）处理大鼠支持细胞或支持细胞 - 生殖细胞共培养物24小时。细胞形态、生殖细胞脱离（GCD）和乳酸丙酮酸产量被用作体外毒性的敏感效应标志物。形态学上，在暴露于二甲基亚砜、1,3 - DNB或DNT异构体24小时后，支持细胞单层保持完整。在100微摩尔的3,4 - DNT下观察到一些明显的细胞毒性：单层被破坏，支持细胞出现广泛的空泡化。用50微摩尔DNT异构体浓度处理的共培养物与用100微摩尔1,3 - DNB处理的细胞非常相似。GCD以剂量依赖性方式增加（0.01和10微摩尔DNT异构体），比对照增加2至10倍。乳酸和丙酮酸产量均随DNT异构体浓度的升高而增加。最敏感的效应出现在3,4 - DNT（10至25微摩尔）。就2,6 - DNT而言，尽管GCD和乳酸产量增加，但仅丙酮酸有最小程度的增加。总体而言，乳酸与丙酮酸产量的比率随DNT剂量的增加而下降。这些结果表明，DNT的四种异构体直接影响支持细胞的形态和功能，其效应与支持细胞毒物1,3 - DNB所见的效应相当。此外，数据支持DNT可能是一种支持细胞毒物的假设。
