从大脑皮质ATP/P2X受体途径探讨PCPA所致失眠的分子机制及酸枣仁汤的干预研究

失眠症是一种影响健康最常见的疾病。ATP及嘌呤P2受体在睡眠调控过程中起到了重要作用。目前认为，神经胶质细胞可释放ATP等递质介导神经系统的信息传递，ATP可作用于嘌呤P2受体，激活不同的细胞内信号转导通路，进而调节睡眠活动。PCPA失眠动物模型是常用的睡眠药物研究的载体，前期研究发现PCPA失眠与神经胶质细胞激活有密切关系，但PCPA失眠是否导致大脑皮质ATP含量变化并将该作用引向何方尚不清楚。本研究拟通过建立PCPA失眠模型，运用高效液相色谱法、免疫荧光双染法、激光共聚焦、Western blotting和RT-PCR等方法，观察PCPA失眠模型中ATP、嘌呤P2X受体及受体后信号活动变化等的影响。本研究将阐明PCPA所致失眠的信号传导机制，揭示酸枣仁汤治疗失眠症的潜在靶点，有利于进一步明确促眠药物作用的环节、作用的靶点，为促眠药物的研发提供分子生物学证据，为失眠症的防治提供新的思路。

Insomnia is one of the most common diseases affecting health. Glial cells play an important role in the processes of sleep regulation; current research shows that ATP is a major astrocytic gliotransmitter in vivo; it is a significant source of extracellular adenosine in the brain; and it can act on purinergic receptors by activating different intracellular signal transduction pathways, thereby regulating sleep activity. The PCPA insomnia animal model is commonly used in insomnia drug research; preliminary studies find that PCPA insomnia and glial cell activation are closely related, but whether PCPA insomnia causes changes in the cerebral cortex ATP level, and the concomitant effects of such changes are unclear. This study uses the established PCPA insomnia model, high performance liquid chromatography and double immunofluorescence staining, confocal laser scanning microscopy, and Western blotting and RT-PCR methods to observe ATP concentration in the cerebral cortex, and the effects of PCPA and Suanzaoren decoction on purine receptors and post-receptor signaling pathways. This study will clarify the signal transduction mechanisms in PCPA insomnia, revealing potential targets of Suanzaoren decoction in the treatment of insomnia. It seeks to further clarify the link targets of sleep drug action, provided molecular evidence to promote sleep medicine research and development, and to provide new ideas for the prevention and treatment of insomnia.