囊泡靶向通路介导的自噬在脊髓损伤后锌毒性中的保护作用及机制研究

项目名称： 囊泡靶向通路介导的自噬在脊髓损伤后锌毒性中的保护作用及机制研究

项目编号： No.81471854

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 梅晰凡

作者单位： 锦州医科大学

项目金额： 80万元

中文摘要： 锌由锌转运体1（ZnT1）介导可明显促进脊髓损伤(SCI)后脑源性神经营养因子(BDNF)分泌，促进SCI后神经修复，但锌过量会导致锌毒性，而机体对锌毒性的保护机制尚未阐明。我们发现在SCI后，损伤脊髓中锌大量聚集，ZnT1表达升高，自噬功能被明显激活，且自噬相关基因与ZnT1的表达在急性期正相关，提示锌能通过ZnT1诱导自噬，而囊泡靶向通路（Cvt）能提高细胞对高锌的耐受力，所以我们认为Cvt特异性自噬通路可能对SCI后神经细胞锌毒性具有保护作用。基于上述工作基础，本课题拟利用自噬APG9L基因敲除小鼠及其SCI模型，在高锌条件下检测脊髓锌的代谢分布，研究脊髓神经细胞存活与自噬通路变化的关系；利用RNA 干扰和基因转染技术干预体外培养脊髓神经元特异性自噬，阐明高锌条件下锌代谢分布与脊髓神经细胞的生物学功能变化情况，揭示Cvt特异性自噬通路在锌毒性中对神经细胞的保护作用及其机

中文关键词： 脊髓损伤；锌毒性；自噬；囊泡靶向通路；锌离子转运体

英文摘要： Brain-derived neurotrophic factor (BDNF) can significantly promote nerve regeneration after spinal cord injury (SCI), zinc could promote the expression of BDNF and its receptor TrKB after SCI, so the zinc has good potential clinical application as a means of treatment of SCI, but zinc has two sides, zinc overload and zinc toxicity after SCI become an urgent problem. Autophagy is activated and play a protective role after SCI, but the extent of its activation is not enough. Our previous studies have shown: as a channel that transport zinc out of intracellular, zinc transporter 1 (ZnT1) expression significantly increased after SCI, and there is a positive correlation between ZnT1 and autophagy in the acute phase, while zinc can induced autophagy, and cytoplasm to vacuole targeting pathway could improve the tolerance to high concentrations of zinc, all these suggested that cytoplasm to vacuole targeting pathway (Cvt) induced autophagy may play a protective role in zinc overload, zinc toxicity after SCI. This project intends to use the autophagy gene knockout mice and their SCI model; using RNA interference and transfection techniques to inhibit or promote autophagy in the cultured neurons; using cell SCI model to detect the protective roles of autophagy under a high level of zinc of in nerve cells in zinc overload and zinc toxicity and to find out the mechanism. All these will solve zinc overload and zinc toxicity after SCI.

英文关键词： spinal cord injury;zinc toxicity;autophagy;cytoplasm to vacuole targeting pathway;zinc transport 1