Sholl S A, Orsini M W, Hitchins D J
J Steroid Biochem. 1983 Aug;19(2):1153-61. doi: 10.1016/0022-4731(83)90410-7.
The steroidogenic potential of hamster tissues, just prior to implantation of the blastocyst in the uterus, was characterized by incubating blastocysts (14) and pieces of endometrium with [1, 2-3H]-androstenedione for 24 h. [3H]-2-Methoxyestradiol was synthesized, but intermediate estrogens were not found. To obtain a more quantitative assessment and comparison of steroidogenic activity, especially aromatase activity, in these tissues as well as in the uterine myometrium and liver and to increase the possibility of recovering estradiol, microsomes were isolated from 244 blastocysts and portions of the other tissues. Microsomes were incubated with [1 alpha, 2 alpha-3H]-testosterone plus [1 beta,2 beta-3H]-testosterone for 6 h. During this time [3H]-metabolites were synthesized by all tissues as indicated by HPLC. [3H]-Androstenedione was noted and values were higher than control levels (medium alone or microsomes from uterine flush fluid) in all samples but liver. [3H]-Estradiol was detected at an elevated level only in the blastocyst sample; however, addition of unlabeled estradiol during the subsequent incubation of endometrial, myometrial and liver microsomes increased the recovery of [3H]-estradiol. Identities of [3H]-2-methoxyestradiol from the first experiment and [3H]-androstenedione and [3H]-estradiol from the second experiment were confirmed by recrystallization. The formation of 3H2O from [beta-3H]-testosterone was used as an index of aromatase activity. After subtracting control medium values, blastocysts were 24-fold more active (dpm/microgram protein) than the endometrium and myometrium in synthesizing 3H2O. While there was no difference in synthetic potential between endometrium and myometrium, aromatase activity in these tissues was greater than that of the liver. Microsomes from uterine flush fluid displayed no capacity for synthesizing 3H2O indicating that the elevated blastocyst levels were not caused by contaminating endometrial cells. These results indicate that all of the tissues examined have the capacity to metabolize C19-steroids to a variety of hormones, including estrogens, and further, that estrogen metabolism occurs rapidly in these tissues. This capacity may be important for providing a suitable hormonal milieu at the time of implantation.
在胚泡植入子宫之前，通过将胚泡（14个）和子宫内膜碎片与[1,2 - 3H]-雄烯二酮一起孵育24小时，来表征仓鼠组织的类固醇生成潜能。合成了[3H]-2-甲氧基雌二醇，但未发现中间雌激素。为了对这些组织以及子宫肌层和肝脏中的类固醇生成活性，尤其是芳香化酶活性进行更定量的评估和比较，并提高回收雌二醇的可能性，从244个胚泡和其他组织的部分中分离出微粒体。将微粒体与[1α,2α-3H]-睾酮加[1β,2β-3H]-睾酮一起孵育6小时。在此期间，如高效液相色谱所示，所有组织都合成了[3H]-代谢物。除肝脏外，所有样品中均检测到[3H]-雄烯二酮，其值高于对照水平（仅培养基或子宫冲洗液中的微粒体）。仅在胚泡样品中检测到[3H]-雌二醇水平升高；然而，在随后的子宫内膜、子宫肌层和肝脏微粒体孵育过程中添加未标记的雌二醇，可提高[3H]-雌二醇的回收率。通过重结晶确认了第一个实验中[3H]-2-甲氧基雌二醇以及第二个实验中[3H]-雄烯二酮和[3H]-雌二醇的同一性。由[β-3H]-睾酮形成3H2O用作芳香化酶活性的指标。扣除对照培养基值后，胚泡在合成3H2O方面的活性（dpm/微克蛋白质）比子宫内膜和子宫肌层高24倍。虽然子宫内膜和子宫肌层之间的合成潜能没有差异，但这些组织中的芳香化酶活性高于肝脏。子宫冲洗液中的微粒体没有合成3H2O的能力，这表明胚泡水平升高不是由污染的子宫内膜细胞引起的。这些结果表明，所有检测的组织都有能力将C19-类固醇代谢为多种激素，包括雌激素，而且，雌激素代谢在这些组织中迅速发生。这种能力对于在植入时提供合适的激素环境可能很重要。
