从PI3K/AKT/IKKβ/NF-κB信号通路探讨电针调控神经免疫功能改善自闭症大鼠行为活动的机制

Synaptic changes and neural immune abnormalities both in autism. The preliminary research results show that the electroacupuncture can improve autism learning and memory of rats with autism and other activities, its mechanism may be related to the regulation in the rat hippocampus synaptic plasticity. Electroacupuncture also have a certain influence of rat hippocampal gliocyte. whether electroacupuncture through regulating nerve immune effect on the behavior of rats with autism activities is not yet clear. This research selects autism rat models for object, observes electroacupuncture for behavior activity changes by open field experiment and water maze rats, and observes electroacupuncture for autism in rats of the influence of immune function by Experiment enzyme-linked immunosorbent (ELISA) method for IL-1β、IL-6 and mmunohistochemical technology for rat hippocampus IBA1 、GFAP protein expression. At the same time , discuss the effect of autism in the rat hippocampus PI3K/AKT/IKKβ/NF-κB pathway by using immunohistochemistry and Western Blot technique observe rat hippocampus PI3K、AKT、IKKβ and NF-κB protein expression, at last verify the working hypothesis of electroacupuncture through regulating neural immune effect on the behavior of rats with autism activities by PI3K/AKT/IKKβ/NF-κB pathway.

突触改变与神经免疫异常并存于自闭症。前期研究表明电针可改善自闭症大鼠学习记忆等行为活动，其机制与电针调控突触可塑性相关；且电针对自闭症大鼠海马胶质细胞存在影响。然而电针是否通过调控神经免疫功能对自闭症大鼠的行为活动产生影响尚未明确。本课题以自闭症大鼠为研究对象，采用旷场实验和水迷宫实验观察电针后自闭症大鼠行为活动的变化；采用ELISA法观察电针后大鼠血清IL-1β和IL-6的变化，采用免疫组织化学技术观察电针后大鼠海马区IBA1和 GFAP蛋白的变化，观察电针对自闭症大鼠神经免疫功能的影响；运用免疫组织化学和Western Blot技术观察电针后大鼠海马区PI3K、AKT、IKKβ和NF-κB蛋白的表达情况，探讨电针对PI3K/AKT/IKKβ/NF-κB通路的影响。综合以上研究验证“电针通过PI3K/AKT/IKKβ/NF-κB信号通路调控神经免疫功能改善自闭症大鼠行为活动”的工作假说。

针刺疗法治疗自闭症疗效确切，已广泛应用于自闭症治疗。研究表明针刺可改善自闭症大鼠的突触功能，然而针刺是否通过调控神经免疫对自闭症大鼠的行为活动产生作用尚未明确。近来神经免疫与自闭症的关系备受关注，本研究以自闭症大鼠为研究对象，观察针刺前后大鼠海马区PI3K、AKT、IKKβ、NF-κB、IBA1和GFAP等蛋白的表达情况，探讨针刺对自闭症大鼠海马PI3K-AKT-IKKβ-NF-κB通路的影响，观察针刺前后自闭症大鼠血清IL-1β、IL-6和IL-10的变化，以验证“针刺通过PI3K-AKT-IKKβ-NF-κB信号通路调控神经免疫功能改善自闭症大鼠行为活动”的工作假说。此外，观察针刺长强穴对自闭症大鼠海马BDNF蛋白表达的影响；并针对近年来临床应用针刺疗法治疗自闭症做一综述，以期为临床针刺治疗自闭症提供参考依据；凝练本课题的针刺操作取穴思路和操作方法，进一步总结小儿针刺的取穴及操作特点，提出针刺治疗小儿疾病（脑瘫）的一些治疗思路。研究结果表明：（1）针刺长强穴对自闭症大鼠行为活动产生影响。（2）针刺长强穴可下调自闭症大鼠海马区NF-κB蛋白的表达，对 PI3K-AKT-IKKβ-NF-κB通路产生一定的影响。（3）针刺长强穴可提高自闭症大鼠血清IL-1β、IL-10水平，对血清IL-6无明显改变。（4）针刺长强穴可上调自闭症大鼠海马BDNF蛋白的表达。