Abstract：
This study aimed to investigate the toxicity and endocrine disrupting potential of a complex mixture of polycyclic aromatic hydrocarbons (PAHs) in the estrogen pathway using hepatocytes of Nile tilapia Oreochromis niloticus, the hepatocytes were exposed to various concentrations of the PAH mixture, and multiple endpoints were evaluated to assess their effects on cell viability, gene expression, oxidative stress markers, and efflux activity. The results revealed that the PAH mixture had limited effects on hepatocyte metabolism and cell adhesion, as indicated by the non-significant changes observed in MTT metabolism, neutral red retention, and crystal violet staining. However, significant alterations were observed in the expression of genes related to the estrogen pathway. Specifically, vitellogenin (vtg) exhibited a substantial increase of approximately 120% compared to the control group. Similarly, estrogen receptor 2 (esr2) showed a significant upregulation of approximately 90%. In contrast, no significant differences were observed in the expression of estrogen receptor 1 (esr1) and the G protein-coupled estrogen receptor 1 (gper1). Furthermore, the PAH mixture elicited complex responses in oxidative stress markers. While reactive oxygen species (ROS) and reactive nitrogen species (RNS) levels remained unchanged, the activity of catalase (Cat) was significantly reduced, whereas superoxide dismutase (Sod) activity, glutathione S-transferase (Gst) activity, and non-protein thiols levels were significantly elevated. In addition, the PAH mixture significantly influenced efflux activity, as evidenced by the increased efflux of rhodamine and calcein, indicating alterations in multixenobiotic resistance (MXR)-associated proteins. Overall, these findings, associated with bioinformatic analysis, highlight the potential of the PAH mixture to modulate the estrogen pathway and induce oxidative stress in O. niloticus hepatocytes. Understanding the mechanisms underlying these effects is crucial for assessing the ecological risks of PAH exposure and developing appropriate strategies to mitigate their adverse impacts on aquatic organisms.
摘要：
本研究旨在研究使用尼罗罗非鱼奥罗罗非鱼肝细胞的雌激素途径中的多环芳烃（PAHs）复杂混合物的毒性和内分泌干扰潜力，肝细胞暴露于各种浓度的PAH混合物，并评估多个终点以评估其对细胞活力的影响，基因表达，氧化应激标志物，和外排活动。结果表明，PAH混合物对肝细胞代谢和细胞粘附的影响有限，如在MTT代谢中观察到的非显著变化所示，中性红保留,和结晶紫染色。然而,在与雌激素途径相关的基因的表达中观察到显著的改变。具体来说,与对照组相比，卵黄蛋白原（vtg）显示出约120％的显着增加。同样，雌激素受体2（esr2）显示出约90％的显着上调。相比之下,雌激素受体1（esr1）和G蛋白偶联雌激素受体1（gper1）的表达没有显着差异。此外，PAH混合物在氧化应激标志物中引起复杂的反应。虽然活性氧（ROS）和活性氮（RNS）水平保持不变，过氧化氢酶（Cat）的活性显着降低，而超氧化物歧化酶(Sod)活性，谷胱甘肽S-转移酶（Gst）活性，和非蛋白硫醇水平显著升高。此外,PAH混合物显著影响外排活性，罗丹明和钙黄绿素的外排增加证明，表明多异种生物抗性（MXR）相关蛋白的改变。总的来说,这些发现,与生物信息学分析相关，强调PAH混合物调节雌激素途径和诱导O.niloticus肝细胞氧化应激的潜力。了解这些影响的潜在机制对于评估PAH暴露的生态风险和制定适当的策略以减轻其对水生生物的不利影响至关重要。
