Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used pharmaceuticals to treat pain, fever and inflammation. NSAIDs are also known to have many side effects including adverse effects on reproduction in both humans and animals. As NSAIDs usage is not regulated they are frequently detected at high concentrations in the environment. In order to understand the effect of NSAIDs on zebrafish sex differentiation, we used seven different NSAIDs which were either Cox-1 selective, Cox-1 biased, non-selective or COX-2 selective. We show that at higher concentration, NSAIDs are toxic to zebrafish embryo as they lead to mortality and hatching delay. Gene expression analysis following short term exposure of NSAIDs led to downregulation of female specific genes including zp2, vtg2 foxl2 and wnt4. Long term exposure of larvae to environmentally relevant concentrations of Cox-2 selective and non-selective NSAIDs resulted in male-biased sex ratio which confirmed the qRT-PCR analysis. However, the Cox-1 selective acetylsalicylic acid and the Cox-1 biased ketoprofen did not alter sex ratio. The observed male-biased sex ratio could also be due to induction of apoptosis process as the genes including p21 and casp8 were significantly upregulated following exposure to the Cox-2 selective and the non-selective NSAIDs. The present study indicates that NSAIDs alter sex differentiation in zebrafish, primarily through inhibition of Cox-2. This study clearly demonstrates that the use of NSAIDs and their release into the aquatic environment should be carefully monitored to avoid adverse effects to the aquatic organisms.

中文翻译：

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非甾体类抗炎药（NSAIDs）在斑马鱼中引起男性偏向的性别分化。

非甾体抗炎药（NSAIDs）是广泛用于治疗疼痛，发烧和炎症的药物。还已知NSAID具有许多副作用，包括对人和动物繁殖的不利影响。由于未规范NSAID的使用，因此经常在环境中以高浓度检测到它们。为了了解NSAID对斑马鱼性别分化的影响，我们使用了7种不同的NSAID，它们分别是Cox-1选择性，Cox-1偏倚，非选择性或COX-2选择性。我们表明，在较高浓度下，NSAID对斑马鱼胚胎有毒，因为它们会导致死亡和孵化延迟。NSAID短期暴露后的基因表达分析导致女性特异性基因（包括zp2，vtg2 foxl2和wnt4）下调。幼虫长期暴露于环境相关浓度的Cox-2选择性和非选择性NSAIDs会导致男性偏向性别比，从而证实了qRT-PCR分析。但是，Cox-1选择性乙酰水杨酸和Cox-1偏向酮洛芬不会改变性别比。观察到的男性偏向性别比也可能归因于凋亡过程的诱导，因为暴露于Cox-2选择性和非选择性NSAID后，包括p21和casp8在内的基因显着上调。本研究表明，NSAID主要通过抑制Cox-2来改变斑马鱼的性别分化。这项研究清楚地表明，应仔细监测NSAIDs的使用及其向水生环境中的释放，以避免对水生生物的不利影响。