环境相关浓度孕激素联合暴露对斑马鱼性腺分化和发育毒理机制研究

The effect of steroid hormones on aquatic organisms in the environment is the hotspot in the field of environmental science research. However, previous researches concentration on environmental estrogen material. Little studies focused on progestins. Various progestins have been found in sewage and aquatic environments. Progestins pose high bioactivity, and may cause significant adverse effects on aquatic wildlife. Previous studies had showed that levonorgestrel posed androgen effect and affect the gonadal development in aquatic organisms, especially in fish. Our previous study had found dydrogesterone also had strong androgen effect and caused male bias. Some significant pathways were enriched by microarray analysis. But it is weak research on the toxicological mechanism of gonad development. Based on our previous study, the project selected dydrogesterone and levonorgestrel that had androgen effect as the target chemical compound to analyze the toxicological effect of zebrafish to dydrogesterone and levonorgestrel as binary mixtures. Present study is to provide a comprehensive analysis of the gene transcription, protein expression, hormone response and histological of zebrafish to investigate the toxicological mechanism of gonad differentiation and development. The research contributes to the environmental hazard and risk assessment of progestins.

环境中类固醇激素对水生生物的毒性是目前环境科学研究领域的热点问题，但是研究主要集中于雌激素物质，而对孕激素物质毒性研究较少。污水和受纳环境中已经检出多种孕激素。孕激素生物活性强，对野生生物有潜在的毒害效应。有研究报道左炔诺酮具有雄激素效应，影响水生生物特别是鱼类性腺分化和发育。申请人前期发现去氢孕酮也有强烈雄激素效应，可诱导斑马鱼雄性化。并利用基因芯片技术富集到重要的生物学通路。但是孕激素对鱼类性腺发育的毒理机制还不清晰。本项目在前期的基础上选择两种具有雄激素效应的孕激素作为目标化合物，以斑马鱼受体生物，研究环境相关浓度联合暴露效应。从基因转录、蛋白表达、激素应答和组织发育等水平综合分析去氢孕酮和左炔诺酮联合暴露对斑马鱼性腺分化和性腺发育的作用机制。本项目将为为孕激素的生态风险评估提供理论支撑。

孕激素物质已在环境中广泛检出，且检测浓度较高，对水环境中生物有潜在的生态毒理效应。本项目主要研究孕激素左炔诺酮、去氢孕酮和醋酸甲羟孕酮对斑马鱼性腺、大脑和眼睛等不同器官的影响。按计划完成了相关研究内容：1）阐述了左炔诺酮、去氢孕酮单一和联合暴露对斑马鱼性腺分化的毒理效应机制。研究发现左炔诺酮和去氢孕酮通过不同生物通路影响斑马鱼性腺分化，这两种孕激素联合暴露对性腺分化的影响与左炔诺酮类似。2）分析了左炔诺酮和去氢孕酮单一和联合暴露对斑马鱼性腺发育的影响。结果表明左炔诺酮和去氢孕酮均增加了成熟卵子和精子比例，其联合暴露呈增强效应。3）探究了孕激素醋酸甲羟孕酮对斑马鱼性腺分化和发育的影响。醋酸甲羟孕酮可具有雄激素活性，可导致雄鱼增加，影响精巢发育。4）探究了去氢孕酮和醋酸甲羟孕酮对斑马鱼大脑的影响。去氢孕酮和醋酸甲羟孕酮均影响促黄体生成素、促卵泡激素和昼夜节律相关基因转录表达。5）揭示了去氢孕酮和醋酸甲羟孕酮对斑马鱼眼睛的毒理效应机制。去氢孕酮和醋酸甲羟孕酮可通过影响光感蛋白影响光周期通路，阻碍光恢复过程。6）探究了雄激素对斑马鱼性腺分化和发育的影响。雄激素雄烯二酮和1,4-雄烯二酮均有雌激素效应，同时可跨代影响F1代斑马鱼。这些结果为评价类固醇激素生态风险提供了重要依据，研究成果发表SCI收录论文9篇，EI收录论文1篇，中文核心论文1篇，协助培养研究生3名。