TLR4 /ROS信号通路在AMD发病中的作用机制研究

项目名称： TLR4 /ROS信号通路在AMD发病中的作用机制研究

项目编号： No.81200702

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学二处

项目作者： 尹莉莉

作者单位： 上海交通大学

项目金额： 23万元

中文摘要： 最近，本课题组发现循环血液中低密度脂蛋白（LDL）增高能引起SD大鼠视网膜发生年龄相关性黄斑变性（AMD）样改变，但作用机制尚不清楚。研究表明，脂质可通过TLR4受体引起的固有免疫介导的炎症通路介导细胞损伤，而免疫炎症反应与AMD发病密切相关。因此本研究拟在原创性研究的基础上，分别在细胞和整体动物水平采用电镜、免疫荧光、RT-PCR、western blot、RNA干扰、转染等技术和方法：（1）观察LDL及OX-LDL对TLR4-/-及WT小鼠视网膜功能及脉络膜-Bruch-RPE复合体的影响；（2）探讨RPE细胞中LDL及OX-LDL对TLR4的表达变化机制；（3）研究LDL及OX-LDL对RPE细胞中TLR4、Lyn蛋白表达、ROS生成和炎症细胞因子表达的影响。通过上述研究，阐明TLR4 /ROS信号通路在早期AMD发病中作用机制，为将来临床早期预防和治疗AMD提供新的靶点。

中文关键词： Toll 样受体4；氧化低密度脂蛋白；年龄相关性黄斑变性；活性氧；

英文摘要： Our team has previously revealed that elevated circulating low-density lipoproteins (LDL) can lead to lipids deposition and RPE dysfunction, which represent for the early age-related macular degeneration (AMD)-like change. But the exact mechanisms are not definitely known. Recent studies suggest that OX-LDL binds to TLR4 and activate the downstream inflammatory pathway to initiate immuno-inflammatory reaction, which is closely related to AMD. So we will set the previous studies as base and do further studies using various biological technologies such as electron microscope、immunofluorescence staining、RT-PCR、western blot、RNAi and translocation to detect: (1) the influence of LDL and oxidized-LDL (oxLDL) to choroid-Bruch-RPE complex of TLR4-/- mice and WT-mice; (2) the gene expression change of RPE cells treated with LDL and oxLDL; (3) the protein expression and phosphorylation level change of TLR4、Lyn、Nox and also ROS and other cytokines production by RPE cells after treated with LDL and oxLDL. Above all, we will elucidate TLR4 / ROS signal pathway in the early onset of AMD mechanism, and thus provide academic basis for further prevention and treatment for AMD.

英文关键词： Toll-like receptor 4；ox-ldl；age related macular degeneration；reactive oxygen species；