新型纳米材料黑磷的发育毒性及其致毒分子机制研究

As the new 2D nanomaterial, black phosphorus has been widely used in electronic and optoelectronic devices, energy production, biomedical applications, antibacterial material and adsorbent material due to its distinct structure and various excellent physicochemical properties. However, with the rapidly increasing production and application of black phosphorus, the discharge and occurrence in aquatic environment should not be ignored. As there is no any report about developmental toxicity of black phosphorus materials on aquatic organism, it is necessary to investigate their developmental toxicity. The current proposal is committed to investigate developmental toxicity and molecular mechanisms of black phosphorus by using zebrafish. The main research contents are as follows. a) To study effects of black phosphorus with different size on the basic development including hatching, survival and morphology by using zebrafish embryo. b) To study effects of black phosphorus with different size on developmental endpoints and organs by using adult zebrafish. c) To explore the molecular mechanisms by using high-throughput transcriptome sequencing and transgenic zebrafish. d) Moreover, to investigate how the natural organic matter affect toxicity of black phosphorus materials in aquatic vertebrates through altering physicochemical property of black phosphorus materials according to actual aquatic environment. This proposal would provide scientific proof in understanding the developmental toxicity of black phosphorus materials and assessing their healthy risk and bio-safety.

黑磷作为一种新型二维材料，由于其独特的结构及众多优异的物理化学性质，被广泛应用于光电设备、能源、生物医药、抗菌剂、吸附剂制备等领域。然而，黑磷的生产与潜在应用快速增加，其在水环境中的排放和赋存将不容忽视，而目前尚无关于黑磷材料水生生物发育毒性的报道，开展其水生生物发育毒性效应研究刻不容缓。因此，本项目拟采用斑马鱼作为模式动物，探究新型纳米材料黑磷的发育毒性及分子机制：（1）采用斑马鱼胚胎，研究不同尺寸的黑磷材料对斑马鱼基础发育如孵化、存活、发育形态的影响；（2）采用成年斑马鱼，研究不同尺寸的黑磷材料对斑马鱼各发育指标及脏器的影响；（3）利用高通量转录组测序技术和转基因斑马鱼模型，研究其毒性作用机制；（4）根据实际水环境，研究天然有机质如何改变黑磷材料理化性质而影响水生生物发育毒性。本项目的实施可望为黑磷纳米材料的发育毒性效应研究提供新的思路，为其健康风险和生物安全性评价提供科学依据。

黑磷是一种新型二维纳米材料，因其独特的性质广泛用于各领域，与此同时，黑磷在环境中的释放及潜在毒性效应引起了广泛关注。尽管目前针对黑磷的毒性已开展了一些研究，但黑磷的发育毒性效应尚不明确。本项目基于经典模式生物斑马鱼，针对不同尺寸黑磷纳米片的发育毒性效应及其分析机制开展了研究。利用扫描电镜、拉曼成像等技术，发现大尺寸黑磷可附着于卵膜表面，且可穿透卵膜附着于胚胎表面，揭示了黑磷在生物体内的赋存特征；黑磷暴露可导致幼鱼心包水肿、心率下降、血流速度减慢，具有明显的发育毒性；结合转基因斑马鱼模型，发现黑磷可造成心脏肿大、血管发育畸形，明确心血管系统为黑磷毒性作用的靶器官；进一步利用转录组测序技术，揭示黑磷产生毒性作用的分子机制与肌肉发育、氧气传输过程、血管内皮生长因子及丝裂原活化蛋白激酶信号通路相关基因有关；小尺寸黑磷可导致幼鱼卵黄水肿、血管发育异常，其作用机制与核糖体合成过程相关基因有关。以上研究结果首次揭示了黑磷的发育毒性效应，阐明了不同尺寸黑磷的毒性效应及分子机制差异，为评价黑磷的环境健康风险提供了重要的科学数据。