Adams S P, Notides A C
Biochem Pharmacol. 1986 Jul 1;35(13):2171-8. doi: 10.1016/0006-2952(86)90588-5.
Oxidative metabolism of [3H]diethylstilbestrol (DES) and the irreversible binding of reactive [3H]DES metabolites to the macromolecules in kidney slices of Syrian golden hamster were investigated. Non-extractable binding of [3H]DES to kidney macromolecules was observed after incubating hamster kidney slices under aerobic conditions (95% O2/5% CO2), but not under anaerobic conditions (100% nitrogen + 2 mM KCN). A number of oxidative metabolites of [3H]DES were detected in the incubation medium of kidney slices incubated under aerobic, but not anaerobic, conditions. The amount of radiolabeled macromolecules formed in male cortical slices under aerobic conditions increased with time of incubation. At a medium concentration of 50 nM [3H]DES, 0.08 pmole [3H]DES equiv./mg dry weight at 30 min and 0.19 pmole [3H]DES equiv./mg dry weight at 120 min were observed. The amount of irreversible [3H]DES-macromolecular complexes also increased with the concentration of [3H]DES in the incubation medium: 1.59 pmole [3H]DES equiv./mg dry weight was formed with 0.5 microM [3H]DES and 21.89 pmole [3H]DES equiv./mg dry weight was formed with 10 microM [3H]DES. Non-extractable [3H]DES binding was detected in all the subcellular fractions of hamster kidney with the highest amount in the microsomal and soluble fractions, followed by the mitochondrial and nuclear fractions. The macromolecular-[3H]DES complexes were solubilized by proteases but not nucleases, suggesting that [3H]DES irreversible binding is principally to the proteins and not the nucleic acids. The cortex as compared with the medulla of the male hamster kidney displayed a 5-fold greater capacity to irreversibly bind [3H]DES metabolites. The male hamster renal cortex showed a 2- to 3-fold greater capacity to form irreversible macromolecular-[3H]DES complexes than the female hamster renal cortex. These data demonstrate that: (1) renal oxidative metabolism of DES results in [3H]DES metabolites binding irreversibly to macromolecules; and (2) the sex and organ site specificity of the [3H]DES-macromolecular binding corresponds with the sex and organ site specificity of renal tumors of the hamster.
研究了叙利亚金黄地鼠肾切片中[³H]己烯雌酚（DES）的氧化代谢以及活性[³H]DES代谢物与大分子的不可逆结合。在有氧条件（95% O₂/5% CO₂）下孵育地鼠肾切片后，观察到[³H]DES与肾大分子的不可提取结合，但在无氧条件（100%氮气 + 2 mM KCN）下未观察到。在有氧而非无氧条件下孵育的肾切片孵育介质中检测到多种[³H]DES的氧化代谢物。在有氧条件下，雄性皮质切片中形成的放射性标记大分子数量随孵育时间增加。在[³H]DES的中等浓度为50 nM时，30分钟时为0.08皮摩尔[³H]DES当量/毫克干重，120分钟时为0.19皮摩尔[³H]DES当量/毫克干重。不可逆的[³H]DES - 大分子复合物的数量也随孵育介质中[³H]DES的浓度增加而增加：0.5 μM [³H]DES时形成1.59皮摩尔[³H]DES当量/毫克干重，10 μM [³H]DES时形成21.89皮摩尔[³H]DES当量/毫克干重。在仓鼠肾的所有亚细胞组分中均检测到不可提取的[³H]DES结合，其中微粒体和可溶组分中的量最高，其次是线粒体和核组分。大分子 - [³H]DES复合物可被蛋白酶溶解但不能被核酸酶溶解，这表明[³H]DES的不可逆结合主要是与蛋白质而非核酸结合。与雄性地鼠肾髓质相比，皮质不可逆结合[³H]DES代谢物的能力高5倍。雄性地鼠肾皮质形成不可逆大分子 - [³H]DES复合物的能力比雌性地鼠肾皮质高2至3倍。这些数据表明：（1）DES的肾氧化代谢导致[³H]DES代谢物与大分子不可逆结合；（2）[³H]DES - 大分子结合的性别和器官部位特异性与仓鼠肾肿瘤的性别和器官部位特异性相对应。
