c-jun氨基末端激酶信号通路在二乙酰吗啡致神经元凋亡中作用的研究

The heroine is namely the diacetylmorphine, which damages the central nerve cells and results in the diseases of the leukoencephalopathy etc., and can also cause sudden death. At present, as the damage mechanism is not clear, and there is still no effective treatment method, and the cause of death remains uncertain. In our earlier study it has been found that the neuronal apoptosis is one of the important causes of the diacetylmorphine damaging the nerve cells, and the activation of the c-jun amino terminal kinase signal channel is an important channel for the diacetylmorphine inducing neuronal apoptosis. .This project will use and culture the primary cultured cerebellar granule neuron and set up the model of the diacetylmorphine causing neuronal apoptosis, and use the technologies of reverse transcription -polymerase chain reaction（RT-PCR) and Western immunoblotting (Western Blotting) etc., to screen out the proteins which have keying action in neuronal apoptosis from several levels, and make deep study on the mechanism of the c-jun amino terminal kinase signal channel speeding up the neuronal apoptosis, aiming to make clear and definite the key protein molecules which regulate and control the neuronal apoptosis, and test and verify the key protein molecules in the animal models, to establish the new targets of treatment. The project is to reveal the neuron damage mechanism and the cause of death through deep study on the essence of the diacetylmorphine making the neuronal apoptosis, and provide scientific experiment base for searching and establishing the new treatment targets.

海洛因即二乙酰吗啡，其损伤中枢神经细胞导致海绵状白质脑病等疾病，也可引起猝死。目前，由于损伤机制不明确，尚无有效的治疗方法，并且死因不明。我们前期研究发现神经元凋亡是二乙酰吗啡致神经细胞损伤的重要原因之一，c-jun氨基末端激酶信号通路的活化是二乙酰吗啡诱导神经元凋亡的重要通路。.本项目将应用培养原代小脑颗粒神经元并建立二乙酰吗啡致神经元凋亡模型，用逆转录聚合酶链反应（RT-PCR）及Western免疫印迹(Western Blotting)等技术，从多个层次筛选对神经元凋亡起关键作用的蛋白，对c-jun氨基末端激酶信号通路促神经元凋亡的机制进行深入研究，旨在明确调控神经元凋亡的关键蛋白分子，并在动物模型上验证关键蛋白质分子，确立治疗的新靶标。本项目通过深入研究二乙酰吗啡致神经元凋亡的本质，揭示其神经元损伤机制及死亡原因，为寻找并确立新的治疗靶标提供科学实验依据。

目前，毒品滥用成为全球最严重的社会问题之一，我国毒品滥用者已逾百万，其中海洛因成瘾人员约占78．3％。海洛因（化学别名二乙酰吗啡）是阿片类毒品的代表，是当今主要的毒品来源，成瘾快，在体内快速代谢，有较强的依赖性，对人体各系统均可造成不同程度的功能损伤。海洛因的应用会导致多种脑部疾病，根据患者的病因可以分为对海洛因依赖性疾病和海洛因导致的感染性疾病,前者包括了戒断综合征、中毒性脑病等；后者包括脑栓塞、脑血管炎等。目前对于JNK MAPK信号通路在海洛因致神经元损伤的发生、发展过程中的作用具体机制阐述不清，只有少数证据提示C-Jun在神经元细胞损伤起关键作用。以往研究主要都集中与海洛因致神经元细胞损伤形态学观察，一定程度证实了海洛因成瘾致脑损伤的主要形式是细胞凋亡，已有充足的依据来证实，对神经细胞凋亡进行调控可有效地减轻海洛因成瘾导致的脑组织损伤的程度。通过对JNK/C-Jun信号通路的关键基因TNF-a，Rho，C-Abl，P-C-Jun，C-Jun，Cytc，Bax，caspase-8和ATF3调控二乙酰吗啡致神经元细胞凋亡的作用机制进行探讨，同时选择性地阻断、抑制或沉默JNK/C-Jun因子的凋亡信号传递通路关键靶点，对JNK/C-Jun信号转导通路在神经元细胞促凋亡的发生机制方面进行深入地探讨，为临床提出以JNK/C-Jun为靶标治疗二乙酰吗啡产生的脑病提供更可靠、更充分的科学依据。