基于NCOA4介导的铁蛋白自噬/铁死亡途径探讨通督醒神针刺改善MCAO学习记忆障碍的机制研究

缺血性脑卒中后学习记忆障碍严重影响患者日常生活，尚缺乏有效治疗措施。铁死亡是一种新型的神经元死亡方式，与学习记忆密切相关。课题组研究发现：通督醒神针刺可通过调控MCAO大鼠海马自噬网络系统，保护神经元，改善学习记忆。最新研究发现，NCOA4介导的铁蛋白自噬参与了神经元的铁死亡过程。故提出假说：通督醒神针刺改善MCAO大鼠学习记忆障碍的作用机制可能与NCOA4介导的铁蛋白自噬/铁死亡途径有关。拟通过体内实验制作MCAO大鼠学习记忆障碍模型，以通督醒神针刺干预；体外实验培养原代海马神经元，构建OGD/R细胞模型以模拟体外MCAO模型，采用电针血清干预。进而采用透射电镜观察大鼠海马神经元超微结构和自噬体情况，并运用Western Blot、ELISA、免疫共沉淀等分子生物学方法检测大鼠海马神经元铁代谢、抗氧化系统、脂质氧化应激、铁蛋白自噬及NCOA4-铁蛋白共表达等，以验证该假说。

Learning and memory impairment after ischemic stroke seriously affects the daily life of patients, and there is still a lack of effective treatment measures. Ferroptosis is a new type of neuron death, which is closely related to learning and memory. Our research team found that Tongdu Xingshen acupuncture can protect neurons and improve learning and memory by regulating the hippocampal autophagy network system of MCAO rats. The latest study found that NCOA4-mediated autophagy of ferritin is involved in the ferroptosis process of neurons. Therefore, the hypothesis is put forward: the mechanism of Tongdu Xingshen acupuncture in improving learning and memory impairment in MCAO rats may be related to the NCOA4-mediated ferritin autophagy/ferroptosis pathway. It is planned to make the learning and memory impairment model of MCAO rats through in vivo experiment, and use Tongdu Xingshen acupuncture intervention; in vitro experiments to cultivate primary hippocampal neurons, construct OGD/R cell model to simulate the vitro MCAO model, and intervene with electroacupuncture serum. Then, transmission electron microscopy will be used to observe the ultrastructure and autophagosomes of rat hippocampal neurons, and molecular biology methods such as Western Blot, ELISA, and immunoprecipitation will be used to detect rat hippocampal neuron iron metabolism, antioxidant system, and lipid oxidation stress, autophagy of ferritin and co-expression of NCOA4-ferritin to verify this hypothesis.