环境浓度下典型抗癫痫药物对稀有鮈鲫脂质代谢的影响及作用机制

The residues and distribution of pharmaceuticals and personal care products in the environment have caused serious harm to wildlife, which raised wide concern around the world. Antiepileptic drugs are widely distributed and detected frequently with high concentration. Although studies have shown that some drugs have effects of environmental endocrine disruptors, other environmental hazards on wildlife are not documented. Endocrine is closely related to abnormal lipid metabolism, and their specific interaction mechanism are still unclear. Therefore, in this project, typical antiepileptic drugs were selected as goal chemicals, Chinese rare minnow with long cycle life stage were exposed to environmental concentration of antiepileptic drugs. The effects of endocrine disrupting and lipid metabolism were analysied. The composition and abundance of gut microorganisms were analyzed by RNA-seq and then their effects on lipid metabolism were analysied. The interation between endocrine system and lipid metabolism induced by antiepileptic drugs were illustrated by molecular biology techniques and analytical methods, which is helpful to provide methods and technical foundation for safety assessment on pharmaceuticals residues. In addition, they also provides basic data for the management of medicines in the water environment.

环境中人类药物和个人护理品的残留和分布对野生生物造成严重危害，引起世界广泛关注。其中抗癫痫类药物在环境中分布广泛、检出率和检出浓度较高，且部分已确认具有内分泌干扰效应，但对野生生物的其他环境危害尚不明确。脂质代谢异常与内分泌关系密切，其在水生生物体内相互作用机制至今未有报道。因此，本项目以典型抗癫痫类药物为研究对象，选择我国特有实验鱼类稀有鮈鲫，开展环境剂量下长周期多生命阶段（幼鱼和成鱼期等）暴露，确定脂质代谢和内分泌激素相关生物标志物响应；利用高通量测序技术，研究肠道菌群组成和丰度，探索其对脂质代谢的影响；采用新分子生物学手段和分析方法，阐明抗癫痫药物对鱼类脂质代谢紊乱与内分泌激素之间的相互作用关系，为药物残留环境危害评估提供方法和技术基础，同时也为水环境药物管理提供基础数据。

近年来，药品和个人护理品的使用量逐年增加，导致药品和个人护理品通过各种直接或者间接途径进入水环境中，从而在水环境中被频繁检出，此外，大量检出的药品和个人护理品对非靶标水生生物潜在的环境危害引起了广泛关注。. 本研究以中国本土鱼类稀有鮈鲫为研究生物，开展了三种抗癫痫药物（卡马西平、丙戊酸钠和奥卡西平）在环境相关浓度下进行为期三个月的长期慢性暴露，探索了抗癫痫类药物对非靶标鱼类稀有鮈鲫的毒性效应以及潜在作用机制，取得的研究结果主要包括以下两个方面：. 环境相关浓度的卡马西平、丙戊酸钠和奥卡西平通过促进稀有鮈鲫脂肪合成和下调脂肪分解相关酶活，从而导致肝脏中显著的脂滴积累（p < 0.05），并诱导稀有鮈鲫性别差异的脂质代谢紊乱。同时卡马西平、丙戊酸钠和奥卡西平能够引起肠道屏障受损，分泌的粘液显著增加（p < 0.05），且肠道微生物组成发生显著变化（p < 0.05）。. 进一步的机制研究发现，丙戊酸钠和奥卡西平对稀有鮈鲫具有内分泌干扰效应，导致雌鱼血浆中的雌二醇和促卵泡激素水平显著增加，引起雄鱼血浆中促卵泡激素水平显著增加（p < 0.05），导致稀有鮈鲫血浆胆固醇水平升高。100µg/L丙戌酸钠诱导雌鱼肝脏Bile secretion信号通路显著变化，可能引起作为胆汁重要成分的胆汁酸水平的变化，导致脂肪代谢异常，进而产生脂质代谢紊乱，而1µg/L雄鱼肝脏PPAR signaling pathway显著变化，通过改变胆固醇合成与分解、脂肪酸氧化等多种生物学过程，引起脂质代谢失衡。奥卡西平和丙戊酸钠的作用机制有所不同。100µg/L奥卡西平诱导雄鱼肠道菌群变化，可能通过AMPK signaling pathway，进而抑制AMPK 激活蛋白激酶的活性，并抑制外周组织中脂肪酸的氧化，进而诱导肥胖和胰岛素抵抗；100µg/L奥卡西平诱导雌鱼肝脏中Estrogen signaling pathway显著改变，引起雌二醇的显著增加，可能通过ER信号通路能够调控肠道微生物菌群，进而引起脂质代谢异常，后续需要进一步研究验证。