Ex vivo assessment of testicular toxicity induced by carbendazim and iprodione, alone or in a mixture.
Ex vivo assessment of testicular toxicity induced by carbendazim and iprodione, alone or in a mixture.
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多菌灵和异菌脲单独或混合诱导的睾丸毒性的离体评估。
DOI：
10.14573/altex.1601253
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
O. Prat
中科院分区：
文献类型：
--
作者：
C. Pisani;S. Voisin;K. Arafah;P. Durand;M. Perrard;M. Guichaoua;P. Bulet;O. Prat
为了测量两种杀菌剂（多菌灵和异菌脲）单独或混合的睾丸毒性，我们使用了大鼠生精小管离体模型，根据3R规则（替换、减少和细化）大大减少了啮齿动物的使用数量。该模型可以代表青春期，这是内分泌干扰物的关键生命时期。通过转录组和蛋白质组分析对细胞修饰进行了三周的跟踪。开发了一种定量和比较方法来估计每种物质对已知途径的干扰程度。这种途径驱动的分析揭示了所有情况下类固醇生成的强烈改变和减数分裂的损害，尽管两种物质的初始分子事件不同。离体细胞遗传学分析证实，两种杀菌剂都会改变第一次减数分裂前期的进程。此外，与单独测试的杀菌剂相比，两种物质的混合物引发的效应大于其累积效应的总和，并且在较短的暴露时间后会损害未来的精子活力。离体培养与“组学”策略的结合加上生理学检查是单独或混合测试物质睾丸毒性的强大工具组合。特别是，蛋白质组学可以鉴定暴露培养物的分泌蛋白中系统差异表达的蛋白质，例如 FUCO 和 PEBP1，这两种蛋白质分别与精子的活力和受精能力相关。这些蛋白质可能是睾丸功能障碍和不育症的潜在生物标志物。
To measure the testicular toxicity of two fungicides (carbendazim and iprodione), alone or in a mixture, we used a rat ex vivo model of seminiferous tubules, greatly reducing the number of rodents used, in accordance with the 3R rule (Replacement, Reduction, and Refinement). This model allows the representation of puberty, a critical life period with regard to endocrine disruptors. The cellular modifications were followed for three weeks through transcriptomic and proteomic profiling analysis. A quantitative and comparative method was developed to estimate how known pathways were disturbed by each substance. This pathway-driven analysis revealed a strong alteration of steroidogenesis and an impairment of meiosis in all cases, albeit the initial molecular events were different for both substances. The ex vivo cytogenetic analysis confirmed that both fungicides alter the course of the first meiotic prophase. In addition, the mixture of both substances triggered effects greater than the sum of their cumulative effects and compromised future sperm motility after a shorter time of exposure compared with the fungicides tested separately. The alliance of an ex vivo culture with "omics" strategies complemented with a physiological examination is a powerful combination of tools for testing substances, separately or in a mixture, for their testicular toxicity. In particular, proteomics allowed the identification of systematically differentially expressed proteins in the secretomes of exposed cultures, such as FUCO and PEBP1, two proteins linked with the motility and fertilizing ability of spermatozoa, respectively. These proteins may be potential biomarkers of testicular dysfunction and infertility.