基于衰老的细胞模型研究PRDX1介导的信号通路在神经干细胞衰老中的调节作用

项目名称： 基于衰老的细胞模型研究PRDX1介导的信号通路在神经干细胞衰老中的调节作用

项目编号： No.31471029

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 神经、认识与心理学

项目作者： 徐俊

作者单位： 同济大学

项目金额： 82万元

中文摘要： 以往的研究显示神经干细胞的老化对神经系统的衰老起着重要作用。本研究拟通过优化羟基脲处理条件，诱导产生持续DNA损伤，通过SA-β-gal染色， 神经干细胞增殖分化能力，其在 G0/G1 细胞周期停滞状态，活性氧自由基的水平及细胞死亡的状况等指标评价神经干细胞的老化状态， 同时检测那些诱发衰老的关键基因(如p16,p21,p53)，以及DNA损伤修复及应答反应相关的重要蛋白xrcc2/3, ku70等的表达变化情况，从而确定是否成功地建立了神经干细胞衰老的离体细胞模型。在此基础上进行基于质谱的定量蛋白组学分析，筛选出调控神经干细胞衰老的关键因子。我们将以PRDX1为例， 利用条件敲除小鼠等分子遗传学手段，结合离体的衰老模型，首先确定PRDX1与神经干细胞衰老的关系，然后进一步研究PRDX1通过与p38 MAPK磷酸脂酶的相互作用对p38MAPK，p16的调控进而调节神经干细胞衰老的机制。

中文关键词： 神经干细胞；衰老；疾病模型；DNA损伤修复；过氧化物酶蛋白1

英文摘要： Aging refers to the physical and functional decline of the tissues over time which often leads to age-related degenerative diseases. Accumulating evidence implicates that the senescence of neural stem cells (NSCs) is of paramount importance to the aging of central neural system (CNS). However, exploration of the underlying molecular mechanisms has been hindered by the lack of proper aging models to allow the mechanistic examination within a reasonable time window. In the present study, we will optimize a hydroxyurea (HU) treatment protocol to induce sustained DNA damage without substantial apoptosis and aim to effectively induce postnatal subventricle NSCs to undergo cellular senescence. The aging of NSCs will be evaluated by senescence-associated-β-galactosidase (SA-β-gal) staining, their proliferation and differentiation capacity, the state of G0/G1 cell cycle arrest, the level of reactive oxygen species (ROS) level and cell death. We will also examine those classical cell-senescence genes as p16, p21 and p53 as well as the key protein involved in various DNA repair pathways such as xrcc2/3, ku70 et al. By this means, we are expecting to establish an in vitro NSC aging model and allow us to further perform quantitative proteomic analysis and outline the key regulators which may have profound impact on the aging of NSCs. Our preliminary data has implied the pivotal role of PRDX1 in NSC aging. In this study, we will further explore its molecular mechanism in depth by utilizing the conditional KO mouse of PRDX1 as well as our cellular aging model. Through these studies we will demonstrate the broad utilities of a cellular aging model and have a better understanding about the nature of NSC senescence and aging.

英文关键词： neural stem cell;aging;disease model;DNA damage and repair;PRDX1