好忘方通过STIM1-Ca2+/Calmodulin-eEF2信号通路治疗阿尔茨海默病的机制研究

传统中医药对阿尔茨海默病（AD）的改善作用近年来引起广泛关注。我们前期发现，补肾豁痰经典方—好忘方可以较为快速、持久地改善AD模型小鼠的认知功能障碍，促进海马神经元的突触传递功能。机制研究表明，好忘方可以显著下调AD模型小鼠海马钙内流调控关键蛋白-STIM1，并影响下游与钙信号关联的负向调控蛋白质合成的关键因子—磷酸化eEF2。基于此，我们提出科学假说：好忘方通过降低AD相关的STIM1介导的钙信号紊乱，逆转Ca2+/Calmodulin-eEF2信号通路导致的突触蛋白合成降低及突触可塑性损伤，从而改善AD认知功能障碍。本项目拟使用Stim1基因敲除小鼠与5×FAD小鼠杂交，结合动物行为学、分子生物学、电生理、钙成像等手段，深入揭示好忘方通过调控STIM1-Ca2+/Calmodulin-eEF2通路改善AD认知功能障碍的机制，为该方的临床应用及进一步药物研发提供参考。

Recently, the effect of Traditional Chinese medicine on improving Alzheimer's disease (AD) has attracted widespread attention . We found that Haowangfang, the classic prescription for invigorating kidney and eliminating phlegm can improve the cognitive dysfunction of AD model mice quickly and lastingly, and promote the synaptic transmission in hippocampal neurons. Mechanical studies showed that Haowangfang can significantly down-regulate STIM1 protein in the hippocampus of AD model mice, which is the key protein regulating calcium influx. Moreover, it can affect the phosphorylated eEF2, a key downstream factor of calcium signaling which negatively modulates protein synthesis. Based on these data, we proposed the scientific hypothesis: Haowangfang reduces AD-related STIM1-mediated calcium signaling disorder, and reverses the reduced protein synthesis and impaired synaptic plasticity caused by Ca2+/Calmodulin-eEF2 signaling pathway, and then improves AD cognitive dysfunction. In this project, we intends to cross Stim1 gene knockout mice with 5×FAD mice, and use behavior test, molecular biology, electrophysiology, calcium imaging and other methods to reveal the mechanisms of Haowangfang regulating the STIM1-Ca2+/Calmodulin-eEF2 pathway to improve cognitive dysfunction in AD. This project will provide a reference for the clinical application of the prescription and further drug development.