针刺调节海洛因成瘾记忆的行为学证据和多巴胺调控机制研究

Addictive memory induced by drug reward and drug-associated environment and cues plays a critical role in the process of heroin dependence and relapse. Long-term heroin use can not only impair normal memory function, but also form aberrant pathological memories of drug-associated environment and cues, in which the latter induces heroin relapse. We observed that acupuncture can positively modulate addictive memory, and has a good effects on psychological craving induced by heroin. We also found that acupuncture modulated firing activity and relevant dimension of dopamine neurons in the ventral tegmental area.In our previous work, it is indicated that the important neural activity area involved in acupuncture is hypothalamus, and acupuncture can suppress the expression of orexin neurons in lateral hypothalamus in heroin dependent rats. Here, we conduct systematic expreriments on the positive effects of acupuncture on addictive memory in different aspects (including memory retention, memory reconsolidation and memory extinction). We will also point out possible mechanisms of acupuncture on heroin addictive memory by investigating the modulating effects of acupuncture on orexin neurons in the lateral hypothalamus or dopamine neurons in the ventral tegmental area. In addition, we will build temporal-spatial discharge feature of dopamine neuronal assembly, conduct correlation studies between neural electrophysiological features and behavioural changes, and analyse possible electrophysiological temporal-spatial discharge feature when addictive memories are modulated by acupuncture.The techniques such as the multi-electrode array recording in freely moving rats, retrograde tracing miRNA control, immunohistochemistry, and the animal model of heroin self-administration induced by drug -associated environment and cues will be used in present study. This study will be helpful to further explore the core mechanisms of acupuncture on heroin relapse.

毒品奖赏作用与用药环境线索产生的病理性记忆是海洛因依赖及复吸的关键。海洛因成瘾者正常记忆水平下降，成瘾相关记忆过程强化，表现出典型的多环节功能紊乱特点。申请人已观察到针刺能干预海洛因成瘾记忆及调节腹侧被盖区多巴胺神经元集群放电和相关维度，结合我们以往工作海洛因依赖者对针刺的主要神经活动区之一为下丘脑，预实验提示针刺能抑制海洛因成瘾大鼠下丘脑外侧区食欲素阳性神经元表达，本研究提出针刺对食欲素-多巴胺调控是针刺干预海洛因成瘾记忆的可能机制。研究采用海洛因成瘾线索记忆模型系统观察针刺对海洛因成瘾记忆多环节良性调节；应用微电极阵列电生理技术、微透析和靶蛋白抑制等技术，观察食欲素-多巴胺调控在针刺干预海洛因成瘾记忆中的作用；开展针刺神经电生理特征和行为学变化的关联性研究，运用生物信息学中的机器学习理论，建构针刺状态下腹侧被盖区多巴胺神经元集群放电的时空特征，以进一步揭示针刺干预海洛因复吸的核心机制。

毒品奖赏作用与用药环境线索产生的病理性记忆是海洛因依赖及复吸的关键。海洛因成瘾者正常记忆水平下降，成瘾相关记忆过程强化，表现出典型的多环节功能紊乱特点。研究采用海洛因成瘾线索记忆模型系统观察针刺对海洛因成瘾记忆多环节良性调节；应用微电极阵列电生理技术、微透析和靶蛋白抑制等技术，观察食欲素-多巴胺调控在针刺干预海洛因成瘾记忆中的作用；开展针刺神经电生理特征和行为学变化的关联性研究，建构针刺状态下腹侧被盖区多巴胺神经元集群放电的时空特征。研究发现针刺能促进海洛因自身给药大鼠觅药记忆行为的消退；低频电针能有效抑制线索和小剂量海洛因诱导的海洛因觅药记忆的维持；低频电针对海洛因成瘾后受损的正常记忆有明显的改善作用；电针能有效干预吗啡成瘾大鼠行为敏化的诱导。研究表明针刺能多环节干预海洛因成瘾记忆，既能改善受损的正常记忆能力，又能多环节干预成瘾记忆过程，为针刺调节成瘾记忆提供了进一步的行为学实验证据。研究还发现，电针能有效降低成瘾大鼠相关环境线索记忆下丘脑Orexin阳性神经元的表达。针刺联合低剂量Orexin A受体拮抗剂能够有效抑制海洛因成瘾大鼠复吸觅药记忆行为。低频电针能下调海洛因成瘾大鼠线索记忆诱导的腹侧背盖区FosB阳性细胞数和pCREB与TH（多巴胺受体标记物）共表达的细胞数目。低频电针能有效降低成瘾记忆相关环境线索暴露时VTA多巴胺动态释放。上述实验结果初步证实下丘脑外侧区食欲素（LH Orexin）—VTA多巴胺神经元在针刺调节成瘾记忆中起重要的作用。电生理实验发现，成瘾大鼠经条件性线索训练，在线索记忆箱VTA 多巴胺能神经元和前额皮层神经元放电频率明显增加，电针能有效降低成瘾大鼠相关记忆环境线索VTA 多巴胺能神经元和前额皮层神经元放电频率，上述结果初步证明，针刺的良性调节来干预海洛因成瘾记忆的复杂改变，以及下丘脑外侧区食欲素（LH Orexin）—VTA多巴胺神经元在针刺调节成瘾记忆中的作用，为进一步揭示针刺戒毒的核心规律提供了依据。