伏隔核区α-突触核蛋白对恒河猴海洛因强化及复燃行为的影响及其相关机制研究

At present, it is very had to find a safe,effective,ethical，ecnomical method to prevent heroin relapse to solve the international problem of treating drug dependence in the world.This study is based on the theory that theα-synuclein could down-regulate the dopamine rewarding neural system and the release of dopamine. The model of heroin self-infusions will be set up in the rhesus monkeys. Lenti-virus α-synclein and target-specific Lenti-virus siRNA will be constructed then will be injected in the nucleus accumbens in rhesus monkeys by the stereotactic technique. Then the effect of stimulation and silencing α-synuclein expression on heroin reinforcement and relapse in the rhesus monkeys will be investigated. And the dopamine content in the relative nucleus with drug dependence will be tested by microdialysis-high performance liquid chromatogram.The texture paraneters of synaptic interface will be also investigated by electron microscope. In all, the study is to investigate the mechanism of the changes of behavior in heroin dependence induced by α-synuclein in the nucleus accumbens.It will offer a new idea of feasible method to solve the international problems of drug dependence.

探寻一种安全有效、符合伦理、经济的防海洛因复吸方法是解决当前国际上治疗药物成瘾的瓶颈与难点。本项目基于α-突触核蛋白可负性调节神经奖赏通路中多巴胺释放的理论基础，以恒河猴海洛因自身给药为模型，构建α-突触核蛋白慢病毒载体与具有靶向特异性的小干扰RNA慢病毒载体，通过立体定向技术注入猴脑伏隔核区，分别使α-突触核蛋白过表达和沉默，观察其对恒河猴海洛因强化及复燃行为的影响，采用微透析－高效液相观察药物成瘾相关核团多巴胺含量变化，电镜观察相关脑区突触界面结构参数的变化，初步探讨伏隔核区α-突触核蛋白诱导恒河猴海洛因成瘾行为的改变及其相关机制，为解决国际性药物成瘾难题提供一种可行的基因新型治疗思路。

药物成瘾问题已经成为全世界范围内公共卫生领域的重要问题之一，防止成瘾药物戒断后复吸是治疗药物成瘾的最大困难，因此，探寻一种安全有效的防复吸方法是解决当前治疗药物依赖的瓶颈与难点。以往研究证实α-突触核蛋白可对神经递质的释放，囊泡再循环以及突触发生、神经元的可塑性起重要影响作用，可负性调节神经奖赏通路中多巴胺释放。本研究通过成功构建α-突触核蛋白小干扰RNA慢病毒载体，在MRI辅助下通过立体定向技术精确定位恒河猴脑部伏隔核区，将慢性病毒注入伏隔核区沉默α-突触核蛋白表达，使用恒河猴封闭式自身给药实验箱，建立恒河猴可卡因自身给药模型通过Western blot技术观察到α-突触核蛋白表达明显下降，采用微透析-高效液相色谱技术观察到恒河猴药物成瘾相关脑区—前额叶皮质及伏隔核区多巴胺含量明显升高，电镜技术观察到恒河猴伏隔核区神经突触活性区长度变长，致密物质变厚，通过可卡因固定比率自身给药实验以及复燃行为诱导实验，观察到沉默伏隔核区α-突触核蛋白表达可导致恒河猴对可卡因强化及复燃行为能力下降。.本研究结果可为探索阿片类药物依赖的机制开拓新思路，并为寻找有效的预防阿片类药物患者戒断后复吸提供基础实验依据。