Environmental Safety Group, Korea Institute of Science and Technology (KIST) Europe Forschungsgesellschaft mbH, 66123 Saarbruecken, Germany.
College of Pharmacy, Chungnam National University, Daejeon 34134, Korea.
Molecules. 2021 Feb 8;26(4):893. doi: 10.3390/molecules26040893.
Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography-tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.
甾体 5-α 还原酶（5AR）负责将甾体还原为 5-α 还原代谢物，例如将睾酮还原为 5-α 二氢睾酮（DHT）。正在开发一种新的甾体 5AR 抑制不良结局途径（AOP），以阐明 5AR 抑制剂对鱼类雌性生殖的抗雌激素作用（AOP 289）。开发了一种使用化学衍生化和液相色谱-串联质谱法分析 DHT 的灵敏方法。建立了一种基于细胞的 5AR 抑制测定法，该方法利用人细胞系、瞬时过表达系统和鱼类细胞系。两种知名的 5AR 抑制剂非那雄胺和度他雄胺在不同系统中的测定 IC 值相当。然而，度他雄胺在鱼类细胞系中的 IC 值低于人细胞系中的 IC 值。非那雄胺对 RTG-2 细胞系的抑制作用更高。这些结果表明，5AR 抑制在物种间可能因结构特征而异。该测定法具有高灵敏度和重现性，适用于各种内分泌干扰化学物质（EDC）的 5AR 抑制筛选。未来的研究将继续评估 EDC 对 5AR 的定量抑制作用，以比较不同物种中的内分泌干扰途径。
