回医蜜煎菖蒲方对脑缺血大鼠学习记忆功能的作用及海马CaMKⅡ信号通路的影响

Learning and memory disorder caused by stroke is an important factor affecting the quality of life of patients with stroke. In the master of HuiHui Prescription, the recipe of Honey fried calamus was recorded which could make the patients suffered from stroke do speech quickly and increase the remember. It was showed in the previous study that Honey fried calamus performed the significantly clinical curative effect on patients suffered from stroke in improving the cognitive function, but the mechanism remains to be elucidated. SD rats would be used in following study as the object of experiment, and nimodipine as the control drug. The model of middle cerebral artery occlusion model was prepared, and then to study the molecular mechanism of the effect on regulating CaMK Ⅱ" molecular switch " of learning and memory. Hippocampal LTP was detected in order to observe the ability of learning and memory. The method of pathology was used to observe hippocampal neuron structure. Syn, MAP-2 and PSD-95 would be detected to observe the changes of synaptic remodeling. NR1 and NR2, GluR1 and GluR2 positive cells of hippocampal neuron would be detected by the method of immunofluorescence labeling in order to observe the wake variation of silent synapses. The technique of molecular biology would be used to detect the expression of CaMKⅡmRNA and the activity of ERK2 and CREB which was the downstream signal molecules of CaMKⅡ, and then to use signal blocker KN-93 in order to observe the change of characteristics of CaMK signal pathway. The mechanisms of Honey Fried Calamus in promoting the restoration of learning and memory function by the effect on the variations of the regulation of cerebral ischemia CaMK II signal pathway in hippocampal LTP after cerebral ischemia, and then the basis would be provided for the further research and application.

中风引起的学习记忆障碍是影响患者生存质量的重要因素，《回回药方》记载蜜煎菖蒲具有使患者“能言快记”的作用，前期临床研究发现，其改善中风认知功能的疗效明显，但作用机制有待阐明。本课题拟以SD大鼠为研究对象，尼莫地平为对照，制备大脑中动脉阻塞模型，从对调控学习记忆的CaMKⅡ“分子开关”的影响研究其分子机制。检测海马LTP以观察学习记忆能力；病理学技术观察海马神经元结构；检测Syn、MAP-2和PSD-95以观察突触重塑变化；免疫荧光标记海马神经元NR1和NR2、GluR1和GluR2阳性细胞，以观察沉默突触唤醒变化；分子生物学技术检测CaMKⅡmRNA及其下游信号分子ERK2和CREB活性，脑内应用信号阻断剂KN-93，观察CaMKⅡ信号通路的变化特点。从蜜煎菖蒲对上述变化的影响揭示其调控脑缺血CaMKⅡ信号通路以增强海马LTP和促进学习记忆功能修复的作用机制，为进一步的研究和应用提供依据。

中风引起的学习记忆障碍是影响患者生存质量的重要因素，《回回药方》记载蜜煎菖蒲具有使患者“能言快记”的作用，本课题以SD大鼠为研究对象，尼莫地平、银杏叶为对照，制备大脑中动脉阻塞模型，病理学技术观察海马神经元结构；检测Syn、MAP-2和PSD-95以观察突触重塑变化；免疫荧光标记海马神经元NR1和NR2、GluR1和GluR2阳性细胞，研究发现中高剂量的蜜煎菖蒲方可显著改善脑缺血再灌注损伤大鼠神经功能和学习记忆能力，对脑缺血再灌注损伤大鼠神经元损伤具有保护作用，蜜煎菖蒲方可促进大鼠脑缺血再灌注损伤后海马神经元突触重塑相关蛋白Syn、MAP-2和PSD-95的表达唤醒沉默突触，其保护海马神经元、改善大鼠神经功能及学习记忆能力的机制可能与促进海马神经元突触重塑有关，为进一步的研究和应用提供依据。