酒精对氯胺酮神经毒性的影响及CREB信号通路相关调控机制研究

Ketamine is a very common anaesthetic in clinic. However, abuse of ketamine becomes more and more serious. As a Party drug (Special K), ketamine is often used in combination with alcohol, which will cause more serious neurotoxicity. However, the characteristics and mechanisms about ketamine combined with alcohol abuse are still not well known and needed to be explored. This research is based on our previous studies which have proved that ketamine can induce neurotoxicity in vivo and in vitro. In this study, characteristics of ketamine combined with alcohol abuse induced neurotoxicity (in vivo and in vitro) will be further studied by behavior tests, high performance liquid chromatography, immunohistochemistry, electron microscope, laser confolcal microscopy, flow cytometry, TUNEL, and Western blot. Involvement of CREB signal pathway will be further explored by comparing ketamine incombination with alcohol treatment and ketamine or alcohol treatment alone. Furthermore, 2-PMPA, a NAAG peptidase inhibitor, will be used to study if it can reduce the neurotoxicity induced by ketamine in combination with alcohol. Its related mechanisms will also be explored. This study will provide useful information for rational use of ketamine in clinic and important experimental data for prevention and treatment of neurotoxicity of ketamine in combination with alcohol abuse.

氯胺酮是临床常用的麻醉药，但滥用的现象愈发严重，作为“舞会药”（K粉）常与酒精联合滥用，引起严重的神经毒性，但其具体的特点和确切的分子机制尚未阐明，是亟待探究的领域。本课题在前期工作证明氯胺酮能够引起体内和体外神经毒性的基础上，以氯胺酮与酒精慢性联合应用诱导的神经毒性（在体、离体）为研究对象，采用行为学、高效液相色谱法、免疫组化、电镜、激光共聚焦、流式细胞术、TUNEL和Western blot等方法研究酒精对氯胺酮神经毒性的影响及其具体特点，与氯胺酮或酒精单独应用进行比较，探索与神经毒性密切相关的CREB信号通路在此过程中的变化规律，进行深入的机制分析。同时考察NAAG肽酶抑制剂2-PMPA对酒精与氯胺酮联合应用所致神经毒性的影响，并分析可能的作用机制。本研究将为氯胺酮临床合理用药，预防和治疗其与酒精联合滥用所致的神经毒性提供重要的实验依据。

氯胺酮是临床常用的麻醉药，但滥用的现象愈发严重，作为“舞会药”（K粉）常与酒精联合滥用，引起严重的神经毒性，但其具体的特点和确切的分子机制尚未阐明，是亟待探究的领域。本课题在前期工作证明氯胺酮能够引起体内和体外神经毒性的基础上，以氯胺酮与酒精慢性联合应用诱导的神经毒性模型（在体、离体）为研究对象，采用行为学、免疫荧光法、电镜、激光共聚焦、流式细胞术和Western blot等方法研究酒精对氯胺酮神经毒性的影响及其具体特点。并与氯胺酮或酒精单独应用进行比较，探索与神经毒性密切相关的CREB信号通路在此过程中的变化规律，进行深入的机制分析。同时考察NAAG肽酶抑制剂2-PMPA对酒精与氯胺酮联合应用所致神经毒性的影响，并分析可能的作用机制。结果表明，氯胺酮与酒精联用能明显增加大鼠的自主活动、刻板行为和共济失调，大脑皮层与海马脑区的细胞形态发生改变、海马区线粒体出现明显的损伤，大脑皮层与海马的多巴胺和谷氨酸含量明显升高，同时CREB信号通路相关蛋白表达明显下调。体外研究采用原代神经细胞培养和PC12细胞进一步证明了上述的现象，且AMPA/KA受体抑制剂CNQX可明显改善二者联用所产生的神经毒性。2-PMPA在胶质细胞存在时也能显著改善二者联用所致的细胞活力降低。本研究将为氯胺酮临床合理用药，预防和治疗其与酒精联合滥用所致的神经毒性提供重要的实验依据。