State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Toxicol Appl Pharmacol. 2010 Sep 15;247(3):222-8. doi: 10.1016/j.taap.2010.06.016. Epub 2010 Jul 6.
Previous studies have demonstrated that perfluorinated chemicals (PFCs) can affect reproduction by disruption of steroidogenesis in experimental animals. However, the underlying mechanism(s) of this disruption remain unknown. Here we investigated the effects and mechanisms of action of 1H, 1H, 2H, 2H-perfluoro-decan-1-ol (8:2 FTOH) on steroidogenesis using a human adrenocortical carcinoma cell line (H295R) as a model. H295R cells were exposed to 0, 7.4, 22.2 or 66.6 microM 8:2 FTOH for 24h and productions of progesterone, 17alpha-OH-progesterone, androstenedione, testosterone, deoxycorticosterone, corticosterone and cortisol were quantified by HPLC-MS/MS. With the exception of progesterone, 8:2 FTOH treatment significantly decreased production of all hormones in the high dose group. Exposure to 8:2 FTOH significantly down-regulated cAMP-dependent mRNA expression and protein abundance of several key steroidogenic enzymes, including StAR, CYP11A, CYP11B1, CYP11B2, CYP17 and CYP21. Furthermore, a dose-dependent decrease of cellular cAMP levels was observed in H295R cells exposed to 8:2 FTOH. The observed responses are consistent with reduced cellular cAMP levels. Exposure to 8:2 FTOH resulted in significantly less basal (+GTP) and isoproterenol-stimulated adenylate cyclase activities, but affected neither total cellular ATP level nor basal (-GTP) or NaF-stimulated adenylate cyclase activities, suggesting that inhibition of steroidogenesis may be due to an alteration in membrane properties. Metabolites of 8:2 FTOH were not detected by HPLC-MS/MS, suggesting that 8:2 FTOH was not metabolized by H295R cells. Overall, the results show that 8:2 FTOH may inhibit steroidogenesis by disrupting the cAMP signalling cascade.
先前的研究表明，全氟化学品（PFCs）可以通过破坏实验动物的类固醇生成来影响生殖。然而，这种破坏的潜在机制尚不清楚。在这里，我们使用人肾上腺皮质癌细胞系（H295R）作为模型，研究了 1H、1H、2H、2H-全氟-癸-1-醇（8:2 FTOH）对类固醇生成的影响及其作用机制。将 H295R 细胞暴露于 0、7.4、22.2 或 66.6μM 8:2 FTOH 24 小时，并用 HPLC-MS/MS 定量测定孕酮、17α-OH-孕酮、雄烯二酮、睾酮、脱氧皮质酮、皮质酮和皮质醇的产生。除了孕酮，8:2 FTOH 处理在高剂量组中显著降低了所有激素的产生。暴露于 8:2 FTOH 显著下调了几种关键类固醇生成酶的 cAMP 依赖性 mRNA 表达和蛋白丰度，包括 StAR、CYP11A、CYP11B1、CYP11B2、CYP17 和 CYP21。此外，在暴露于 8:2 FTOH 的 H295R 细胞中观察到细胞内 cAMP 水平呈剂量依赖性下降。观察到的反应与细胞内 cAMP 水平降低一致。暴露于 8:2 FTOH 导致基础（+GTP）和异丙肾上腺素刺激的腺苷酸环化酶活性显著降低，但不影响总细胞 ATP 水平或基础（-GTP）或 NaF 刺激的腺苷酸环化酶活性，表明类固醇生成的抑制可能是由于膜性质的改变。HPLC-MS/MS 未检测到 8:2 FTOH 的代谢物，表明 8:2 FTOH 未被 H295R 细胞代谢。总的来说，这些结果表明，8:2 FTOH 可能通过破坏 cAMP 信号级联来抑制类固醇生成。
