Diclofenac (DCF) and caffeine (CAF) are persistent pharmaceuticals that occur in mixtures in the aquatic ecosystems causing effects in the reproductive physiology of aquatic organisms. This study evaluated the physiological reproductive responses of Astyanax altiparanae males exposed to nominal concentrations of DCF (3.08 mg L-1) and CAF (9.59 mg L-1) separately and combined, for 96 h. The steroids profile, estrogenic biomarker vitellogenin (vtgA), testes and liver morphology, and also mortality of males were assessed. DCF and CAF degradation was 5% of the initial concentration for 24 h. The LC50 of the DCF and CAF were 30.8 mg L-1 and 95.9 mg L-1, respectively. Males exposed to DCF and CAF exhibited a reduction of 17β-Estradiol (E2) concentration compared to control (CTL). Similarly, testosterone (T) was also reduced in the DCF treatment, but this response was not observed in 11-Ketotestosterone (11-KT). Males exposed to DCF + CAF combined did not exhibit differences in T, E2 and 11-KT steroids. The vtgA gene expression and the sperm concentration did not change among the treatments. Moreover, acute exposure revealed a hypertrophy of hepatocytes cells in the DCF and DCF + CAF treatments. In conclusion, DCF and CAF, isolated, exhibit an endocrine disruptive activity in A. altiparanae male, an opposite response observed with the mixture of both compounds that abolishes the endocrine disruptive effects. DCF seems to be more toxic for this species, altering also hepatocytes morphology.

中文翻译：

---

双氯芬酸和咖啡因对Astyanax altiparanae雄性（Teleostei：Characiformes：Characidae）的内分泌干扰作用。

双氯芬酸（DCF）和咖啡因（CAF）是持久性药物，它们以混合形式存在于水生生态系统中，对水生生物的生殖生理产生影响。这项研究评估了分别暴露并联合使用96小时的名义浓度的DCF（3.08 mg L-1）和CAF（9.59 mg L-1）的Astyanax altiparanae雄性的生理生殖反应。评估了类固醇概况，雌激素生物标志物卵黄蛋白原（vtgA），睾丸和肝脏形态，以及男性的死亡率。DCF和CAF在24小时内的降解为初始浓度的5％。DCF和CAF的LC50分别为30.8 mg L-1和95.9 mg L-1。与对照（CTL）相比，暴露于DCF和CAF的雄性表现出17β-雌二醇（E2）浓度降低。同样，在DCF治疗中，睾丸激素（T）也降低了，但在11-酮睾酮（11-KT）中未观察到这种反应。暴露于DCF + CAF的男性在T，E2和11-KT类固醇方面无差异。在处理之间，vtgA基因表达和精子浓度没有变化。此外，急性暴露表明在DCF和DCF + CAF治疗中肝细胞肥大。总之，分离的DCF和CAF在拟南芥男性中表现出内分泌破坏活性，这两种化合物的混合物观察到相反的反应，从而消除了内分泌破坏作用。DCF似乎对该物种更具毒性，还会改变肝细胞的形态。在处理之间，vtgA基因表达和精子浓度没有变化。此外，急性暴露表明在DCF和DCF + CAF治疗中肝细胞肥大。总之，分离的DCF和CAF在拟南芥男性中表现出内分泌破坏活性，这两种化合物的混合物观察到相反的反应，从而消除了内分泌破坏作用。DCF似乎对该物种更具毒性，还会改变肝细胞的形态。在处理之间，vtgA基因表达和精子浓度没有变化。此外，急性暴露表明在DCF和DCF + CAF治疗中肝细胞肥大。总之，分离的DCF和CAF在拟南芥男性中表现出内分泌破坏活性，这两种化合物的混合物观察到相反的反应，从而消除了内分泌破坏作用。DCF似乎对该物种更具毒性，还会改变肝细胞的形态。