Zheng Xiangqin, Wei Yuexin, Chen Jiadong, Wang Xia, Li Dinggang, Yu Chengjun, Hong Yifan, Shen Lianju, Long Chunlan, Wei Guanghui, Wu Shengde
Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing 400014, China.
Toxics. 2023 Mar 30;11(4):328. doi: 10.3390/toxics11040328.
Difenoconazole (DFZ) is a broad-spectrum triazole fungicide that is widely utilized in agriculture. Although DFZ has been demonstrated to induce reproductive toxicity in aquatic species, its toxic effects on the mammalian reproductive system have yet to be fully elucidated. In vivo, male mice were administered 0, 20 or 40 mg/kg/d of DFZ via oral gavage for 35 days. Consequently, DFZ significantly decreased testicular organ coefficient, sperm count and testosterone levels, augmented sperm malformation rates, and elicited histopathological alterations in testes. TUNEL assay showed increased apoptosis in testis. Western blotting results suggested abnormally high expression of the sperm meiosis-associated proteins STRA8 and SCP3. The concentrations of retinoic acid (RA), retinaldehyde (RE), and retinol (ROL) were increased in the testicular tissues of DFZ-treated groups. The mRNA expression level of genes implicated in RA synthesis significantly increased while genes involved in RA catabolism significantly decreased. In vitro, DFZ reduced cell viability and increased RA, RE, and ROL levels in GC-2 cells. Transcriptome analysis revealed a significant enrichment of numerous terms associated with the RA pathway and apoptosis. The qPCR experiment verified the transcriptome results. In conclusion, our results indicate that DFZ exposure can disrupt RA signaling pathway homeostasis, and induce testicular injury in mice testes.
苯醚甲环唑（DFZ）是一种广泛应用于农业的广谱三唑类杀菌剂。尽管已证明DFZ会对水生物种产生生殖毒性，但其对哺乳动物生殖系统的毒性作用尚未完全阐明。在体内实验中，雄性小鼠通过灌胃给予0、20或40 mg/kg/d的DFZ，持续35天。结果显示，DFZ显著降低了睾丸脏器系数、精子数量和睾酮水平，提高了精子畸形率，并引发了睾丸的组织病理学改变。TUNEL检测显示睾丸细胞凋亡增加。蛋白质免疫印迹结果表明，精子减数分裂相关蛋白STRA8和SCP3的表达异常升高。在DFZ处理组的睾丸组织中，视黄酸（RA）、视黄醛（RE）和视黄醇（ROL）的浓度升高。参与RA合成的基因的mRNA表达水平显著增加，而参与RA分解代谢的基因则显著降低。在体外实验中，DFZ降低了GC-2细胞的活力，并增加了细胞内RA、RE和ROL的水平。转录组分析显示，与RA途径和细胞凋亡相关的众多术语显著富集。qPCR实验验证了转录组结果。总之，我们的结果表明，接触DFZ会破坏RA信号通路的稳态，并诱导小鼠睾丸损伤。
