内源性大麻素及其1型受体参与电针治疗VPA自闭症模型大鼠的机制研究

More and more people attach importance to autism spectrum disorders because of a huge increase in the rate of diagnosed cases.There is no specific treatment for those problems. Clinical randomized controlled trials and animal model study indicate electroacupuncture could affect ASD-like behaviors. Possible mechanisms were not clear. Our recent study have demonstrated that the therapeutic effect of electroacupuncture on the behavior of the VPA model of autistic rats could be reversed by the cannabinoid 1 receptor antagonist. Based on the previous work, the proposed study will perform a single injection of VPA to pregnant rats on gestational day 12.5 yielded offspring autism rat model. Through the treatment of different frequency electroacupuncture, the expression level of oxytocin and endogenous cannabinoid AEA was detected in the brain reward system, and the key brain region was screened. And through the application of oxytocin receptor specificity antagonists, selective CB1R knockout, those results indicate that endocannabinoids and its type 1 receptor involved in electricacupuncture treatment of VPA model behavioral symptoms,driven by oxytocin.This study, for the first time, discusses the mechanism of electroacupuncture on autism from the perspective of endocannabinoid, and provides theoretical basis for the intervention and treatment of autism behavioral symptoms.

伴随发病率的增高，自闭症越来越多受到人们和社会的重视，目前缺乏有效的治疗。临床随机对照实验及动物模型研究显示电针可以改善自闭症行为学症状，但其机制并不明确。我们前期研究发现：电针治疗可以改善丙戊酸钠（VPA）自闭症模型大鼠行为学症状，且这一治疗作用可以被大麻素1型受体拮抗剂所逆转。在此基础上，本研究以孕鼠孕12.5天腹腔注射VPA制备子代自闭症模型为研究对象，通过不同频率电针治疗，检测大脑奖赏系统相关脑区催产素、内源性大麻素AEA表达水平，筛选关键脑区。并通过应用催产素受体特异性拮抗剂，选择性CB1R基因敲除进行验证，阐述内源性大麻素及其1型受体参与电针治疗VPA自闭症模型行为学症状的催产素机制。本研究首次从内源性大麻素这一角度，探讨电针治疗自闭症的作用机制，为电针作为干预和治疗自闭症行为学症状提供理论依据。

基础和临床研究均已经提示电针可以改善自闭症的核心及伴随症状，但其机制并不明确。本研究在前期工作基础上，首次从内源性大麻素及其1型受体（CB1R）这一角度探讨电针治疗VPA自闭症模型小鼠的可能机制。课题组首先通过使用动物行为学及分子生物学筛选出电针治疗VPA自闭症模型小鼠社交障碍的最佳电针频率是2Hz，内源性大麻素信号系统参与电针治疗VPA自闭症小鼠社交障碍的关键脑区在伏隔核(NAc)。接下来通过系统和局部脑区注射CB1受体拮抗剂AM251阻断电针这一治疗作用。最后通过使用基于CB1R-LOXP小鼠局部脑区注射rAAV-Cre病毒实现特异性敲除NAc中GABA能或谷氨酸能神经元上的CB1受体，三箱社交行为学实验结题提示: 特异性敲除NAc中GABA 能神经元上的CB1受体显著阻断电针治疗VPA自闭症小鼠的社交障碍症状。综上，我们的研究提示：内源性大麻素及其1型受体参与电针治疗VPA自闭症小鼠社交障碍症状，NAc中GABA能神经元上的CB1受体可能是电针治疗的重要靶点。