miR-142-5p/DAX1-dependent regulation of P450c17 contributes to triclosan-mediated testosterone suppression
miR-142-5p/DAX1-dependent regulation of P450c17 contributes to triclosan-mediated testosterone suppression
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DOI：
10.1016/j.scitotenv.2020.137280
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发表时间：
2020-05-15
影响因子：
9.8
通讯作者：
Liu, Changjiang
中科院分区：
文献类型：
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作者：
Duan, Peng;Huang, Xu;Liu, Changjiang
三氯生（TCS）是一种强效的抗菌和抗真菌化合物，广泛用于各种日常用品中。TCS也被认为是一种潜在的内分泌干扰物，具有抗雄激素作用。在我们之前的研究中，我们发现TCS通过miR - 6321/JNK/Nur77级联抑制睾丸类固醇生成，但miR - 142 - 5p和P450c17异常表达在这一分子事件中的作用仍不清楚。因此，为了验证miR - 142 - 5p和P450c17可能在TCS暴露后睾酮下降中以其他方式发挥重要作用的假设，本研究使用了斯普拉格 - 道利大鼠和大鼠睾丸间质细胞系。结果表明，TCS暴露后，睾丸组织形态异常改变，睾酮水平下降。TCS过表达的miR - 142 - 5p直接靶向JAK1/STAT1通路。双向免疫共沉淀（Co - IP）实验以及使用STAT1激活剂表明，STAT1可以与Sp1相互作用并对其进行调节。TCS处理后，DNMT1和DNMT3β的活性、mRNA水平和蛋白质表达均下降。Sp1沉默、染色质免疫沉淀（ChIP）和定量聚合酶链反应（qPCR）实验表明，Sp1通过直接结合到DNMT1的启动子区域来调节DNMT1的表达。尽管DAX1启动子的DNA甲基化状态未受影响，但TCS通过DNMT1诱导DAX1的转录和翻译，进而导致类固醇生成相关的P450c17受到抑制。综上所述，TCS诱导的miR - 142 - 5p通过JAK1/STAT1通路以及下游的Sp1/DNMT1/DAX1级联抑制P450c17，最终促使睾酮水平下降。（C）2020爱思唯尔有限公司。保留所有权利。
Triclosan (TCS) is a potent antibacterial and antifungal compound that is extensively used in various daily products. TCS is also considered as an underlying endocrine disruptor and has anti-androgenic effects. In our previous work, we found that TCS suppressed testicular steroidogenesis via the miR-6321/JNK/Nur77 cascade, but roles of the abnormal expression of miR-142-5p and P450c17 in this molecular event were still unknown. Therefore, to verify the hypothesis that miR-142-5p and P450c17 might significantly function in other manner in testosterone decline after TCS exposure, Sprague-Dawley rats and the rat Leydig cell line were used in this study. Results showed that after TCS exposure, testicular histomorphology was abnormally changed and testosterone level was declined. Overexpressed miR-142-5p by TCS directly targeted the JAK1/STAT1 pathway. Bidirectional Co-IP assays and the use of STAT1 activator demonstrated that STAT1 could interact with and regulate Sp1. The activity, mRNA level, and protein expression of DNMT1 and DNMT3 beta, were all decreased after TCS treatment. Sp1 silencing. ChIP, and qPCR assays showed that Sp1 regulated DNMT1 expressions by directly binding to the promoter region of DNMT1. Though the DNA methylation status of the DAX1 promoter was not affected, TCS induced the transcription and translation of DAX1 by DNMT1, in turn leading to the inhibition of steroidogenic P450c17. Taken together, TCS- induced mi R-142-5p inhibits P450c17 by the JAK1/STAT1 pathway and down- stream Sp1/DNMT1/DAX1 cascade, finally facilitating the decrease in testosterone levels. (C) 2020 Elsevier B.V. All rights reserved.