基于melatonin受体亚型内吞循环异常的丙戊酸分子毒理机制研究

产前服用抗癫痫药物丙戊酸钠增加儿童自闭症风险。前期研究发现丙戊酸钠自闭症模型诱发了神经生物化学及关联功能改变，但其分子毒理学机制并不清楚。本项目以膜表面melatonin受体亚型（MTNR1）为着眼点，采用time lapse及荧光能量共振转移多种分子生物学方法从以下几个方面揭示丙戊酸钠介导的神经毒理学机制：①确认丙戊酸钠是否诱导膜表面melatonin受体亚型内吞循环及信号通路紊乱；②深入研究丙戊酸钠介导melatonin受体亚型内吞循环异常的分子毒理学机制研究；③药物调控/逆转丙戊酸钠介导的melatonin受体亚型内吞循环异常研究。本研究将阐明丙戊酸是如何影响了melatonin/MTNR1系统的紊乱和其具体的分子毒理机制，并寻找可调控/逆转丙戊酸介导MTNR1内吞异常的药物。也将为了解丙戊酸诱导自闭症发生的分子机制奠定夯实的基础，为进一步防治自闭症提供新型药物靶点及治疗策略。

Fetal valproate exposure is associated with increased risks of autism spectrum disorder. Melatonin receptors are G protein-coupled receptors (GPCRs) that mediate the physiological effects of melatonin. The present study was designed to address the mechanistic details of disturbance of melatonin receptor subtype 1 (MTNR1A) internalization upon valproate exposure. By using time-lapse confocal microscopy, we first determine in living cells that valproic acid (VPA) treatment whether interfered with the internalization kinetics of MTNR1A in the presence of melatonin. Furthermore, we examined the potential molecular toxicological mechanism of valproic acid on dynamic changes of MTNR1A intracellular trafficking; the association between proteins will be detected by using the fluorescence resonance energy transfer (FRET) imaging and immunoprecipitation. Finally, we will investigate the effect of pharmacological modulation on the VPA-mediated disturbance of MTNR1A intracellular trafficking and signaling. Our data may provide useful insights into novel therapies that relieve the effect of VPA on intracellular MTNR1A trafficking and signaling may help lower the risk of autism spectrum disorders.