Lv Xuan, Pan Liumeng, Wang Jiaying, Lu Liping, Yan Weilin, Zhu Yanye, Xu Yiwen, Guo Ming, Zhuang Shulin
College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058, China; Key Laboratory of Health Risk Factors for Seafood of Zhejiang Province, Zhoushan 316022, China.
College of Environmental and Resource Sciences, Zhejiang University, Hangzhou 310058, China.
Environ Pollut. 2017 Mar;222:504-512. doi: 10.1016/j.envpol.2016.11.051. Epub 2016 Dec 21.
Triazole fungicides are widely used as broad-spectrum fungicides, non-steroidal antiestrogens and for various industrial applications. Their residues have been frequently detected in multiple environmental and human matrices. The increasingly reported toxicity incidents have led triazole fungicides as emerging contaminants of environmental and public health concern. However, whether triazole fungicides behave as endocrine disruptors by directly mimicking environmental androgens/antiandrogens or exerting potential androgenic disruption indirectly through the inhibition of cytochrome P450 (CYP450) enzyme activity is yet an unresolved question. We herein evaluated five commonly used triazole fungicides including bitertanol, hexaconazole, penconazole, tebuconazole and uniconazole for the androgenic and anti-androgenic activity using two-hybrid recombinant human androgen receptor (AR) yeast bioassay and comparatively evaluated their effects on enzymatic activity of CYP3A4 by P450-Glo™ CYP3A4 bioassay. All five fungicides showed moderate anti-androgenic activity toward human AR with the IC ranging from 9.34 μM to 79.85 μM. The anti-androgenic activity remained no significant change after the metabolism mediated by human liver microsomes. These fungicides significantly inhibited the activity of CYP3A4 at the environmental relevant concentrations and the potency ranks as tebuconazole > uniconazole > hexaconazole > penconazole > bitertanol with the corresponding IC of 0.81 μM, 0.93 μM, 1.27 μM, 2.22 μM, and 2.74 μM, respectively. We found that their anti-androgenic activity and the inhibition potency toward CYP3A4 inhibition was significantly correlated (R between 0.83 and 0.97, p < 0.001). Our results indicated that the risk assessment of triazole pesticides and structurally similar chemicals should fully consider potential androgenic disrupting effects and the influences on the activity of CYP450s.
三唑类杀菌剂被广泛用作广谱杀菌剂、非甾体抗雌激素药物以及用于各种工业应用。它们的残留物在多种环境和人体基质中经常被检测到。越来越多报道的毒性事件使三唑类杀菌剂成为环境和公共卫生关注的新兴污染物。然而，三唑类杀菌剂是通过直接模拟环境雄激素/抗雄激素发挥内分泌干扰作用，还是通过抑制细胞色素P450（CYP450）酶活性间接发挥潜在的雄激素干扰作用，仍是一个未解决的问题。我们在此使用双杂交重组人雄激素受体（AR）酵母生物测定法评估了五种常用的三唑类杀菌剂，包括联苯三唑醇、己唑醇、戊唑醇、戊唑醇和烯效唑的雄激素和抗雄激素活性，并通过P450-Glo™ CYP3A4生物测定法比较评估了它们对CYP3A4酶活性的影响。所有五种杀菌剂对人AR均表现出中等程度的抗雄激素活性，IC范围为9.34 μM至79.85 μM。在人肝微粒体介导的代谢后，抗雄激素活性没有显著变化。这些杀菌剂在环境相关浓度下显著抑制CYP3A4的活性，效力排名为戊唑醇>烯效唑>己唑醇>戊唑醇>联苯三唑醇，相应的IC分别为0.81 μM、0.93 μM、1.27 μM、2.22 μM和2.74 μM。我们发现它们的抗雄激素活性与对CYP3A4抑制的抑制效力显著相关（R在0.83至0.97之间，p < 0.001）。我们的结果表明，三唑类农药和结构相似化学品的风险评估应充分考虑潜在的雄激素干扰效应以及对CYP450s活性的影响。
