养心安神法调控腺苷参与睡眠剥夺模型基底前脑能量代谢重构的机制研究

睡眠剥夺导致能量代谢重构，增加肥胖、胰岛素抵抗、代谢综合征等疾病风险。前期工作发现，养心安神法提高AVP表达，而AVP是AMPK信号通路的上游激活因子。慢性睡眠剥夺引起基底前脑星形胶质细胞释放腺苷，激活A1腺苷受体，养心安神法可通过提高AVP表达，激活AMPK/PPAR/PGC-1α信号通路，调控星形胶质细胞与神经元交流对话，参与调控睡眠觉醒和能量代谢重构。本项目在前期基础上，拟检测睡眠剥夺模型整体效应、代谢生化指标、代谢模式和代谢轮廓变化；GFAP免疫荧光双标法定位AS与Glu、GABA共表达，构建A1ARsiRNA和A1AR过表达质粒转染等研究AS与神经元交流对话机制；在生物信息学筛选SD中能量代谢差异表达基因和关键通路基础上，结合WB、RT-PCR检测AMPK/PPAR/PGC-1α信号通路变化，为睡眠剥夺防治提供新的视角和思路，进一步丰富和阐释养心安神法的科学内涵。

Sleep deprivation leads to the reconstruction of energy metabolism and increases the risk of obesity, insulin resistance, metabolic syndrome and other diseases. In previous work, it was found that the expression of AVP was enhanced by the heart-nourishing and mind-calming therapy, and AVP was the upstream activator of the AMPK signaling pathway. Chronic sleep deprivation induced basal forebrain astrocytes releasing adenosine, which activating A1 adenosine receptors. Heart-nourishing and mind-calming therapy can activate the AMPK/PPAR/PGC-1 signal pathway, which can regulate the astrocytes and neurons crosstalk, and participate in the regulation of sleep wake and energy metabolic remodeling via improve the expression of AVP. Accordingly, the project will observe the overall effect of sleep deprivation, biochemical indicator, metabolic pattern and metabolic profile; localization of AS, Glu, GABA through GFAP immunofluorescent double staining , study the interaction mechanism of AS and neuron through construction of A1ARsiRNA and A1AR overexpressed plasmid; AMPK/PPAR/PGC-1α signaling pathway changes based on bioinformatics screening of energy metabolism differentially expressed genes and key signaling pathways through Western blot, RT-PCR methods.It will provide new perspectives and ideas for prevention and treatment of sleep deprivation, and further enrich the scientific connotation of heart-nourishing and mind-calming therapy.