肝郁脾虚证为临床常见证候，其主症之一为食欲不振。下丘脑nesfatin-1（NES1）-催产素（OT）-脑干前阿黑皮素原（POMC）神经通路在食欲调节过程中起重要作用，NES1为此通路的关键分子。课题组前期实验发现：该证候大鼠模型下丘脑NES1水平明显升高，与进食量的降低呈正相关。本项目提出假说：“肝郁脾虚证食欲不振的生物学基础与下丘脑NES1-OT-脑干POMC食欲神经通路的激活有关”。拟在复制该证候大鼠模型基础上，分别以NES1抗体、OT受体阻滞剂、POMC受体阻滞剂对此神经通路的三个关键节点进行阻断，以抑制该神经通路，采用动物行为学、脑室微量注射法、RT-qPCR、原位杂交、免疫组化、免疫荧光双标、电镜等技术方法，并通过逍遥散以方测证，研究基于下丘脑NES1-OT-脑干POMC食欲神经通路在肝郁脾虚证食欲不振形成的分子机制。这对于深刻理解肝郁脾虚证食欲不振的形成机理具有重要意义。

The syndrome of stagnation of liver qi and spleen deficiency is one of common syndromes in clinic, the main symptoms of which is loss of appetite. It has been discovered that the activation of hypothalamic/nesfatin-1 (NES1)-oxytocin (OT)-brainstem/pro-opiomelanocortin (POMC) neuronal pathway could effectively regulate appetite, in which the suppressor peptide NES1 plays a pivotal role. Our former research indicated that syndrome of stagnation of liver qi and spleen deficiency showed increased NES1 levels, which were positively correlated with the reduction in food intake in rat model. Therefore, in this current study, we propose a hypothesis that the mechanism of loss of appetite of syndrome of stagnation of liver qi and spleen deficiency may be related to the activation of hypothalamic/NES1-OT-brainstem/POMC neuronal pathway. This study firstly develops the rat model of syndrome of stagnation of liver qi and spleen deficiency that will be effectively assessed with the effects of Xiaoyao Powder highly correlating the syndrome and the behavioral experiments. Then, three key nodes in this neural pathway related to appetite are respectively blocked with the NES1 antibody, OT receptor blocker and α-melanocyte stimulating hormone (α-MSH, a POMC derived peptides) receptor blocker via the intracerebroventricular microinjection, and meanwhile the methods including reverse transcription-quantitative polymerase chain reaction, in situ hybridization, immunohistochemistry, double immunofluorescence, electron microscopy are used to detect the expressions of related gene and protein in the neural pathway. Overall, the purpose of this study is to clarify the molecular mechanism of loss of appetite in syndrome of stagnation of liver qi and spleen deficiency based on the hypothalamic/NES1-OT-brainstem/POMC neuronal pathway. This study will be of great significance in understanding the biological basis of loss of appetite manifested in the syndrome of stagnation of liver qi and spleen deficiency.