基于微流控芯片的发育性、变形性大脑疾患的表观遗传学调控机制研究

项目名称： 基于微流控芯片的发育性、变形性大脑疾患的表观遗传学调控机制研究

项目编号： No.61302030

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 无线电电子学、电信技术

项目作者： 李彧媛

作者单位： 大连医科大学

项目金额： 26万元

中文摘要： 脑保护与脑损伤修复研究一直是生物医药研究领域的热点和难点。表观遗传学调控机制在发育性、变性性大脑疾患发展中起着至关重要的作用，且具有可逆性，为疾病治疗提供了乐观的前景。本研究拟采用ELISA分析神经细胞，如毛细管内皮细胞（E）、星形胶质细胞（A）、神经元（N）特异性靶蛋白的表达差异，寻找与大脑疾患相关的蛋白靶分子；采用RT-PCR、Western Blot、免疫荧光细胞化学染色以及甲基化特异性PCR方法体外研究影响各蛋白靶分子表达的表观遗传学调控机制；利用高通量的DNA甲基化微流控芯片分析检测临床样品。为进行体外研究，我们首次利用Transwell将来源于同一种属动物同一部位的E、A、N三种原代细胞进行立体共培养，构建原代同源EAN三细胞四维模型，为机制的体外研究提供更准确可靠的研究载体。本研究将为深入了解发育性、变性性大脑疾患的发病机制和研发新型治疗药物拓展新思路。

中文关键词： 神经炎症；多发性硬化；表观遗传学调控；DNA甲基化；PGE2

英文摘要： The study on neuroprotection and neuroregeneration has been a hotspot and difficulty of biomedical research. In the central nervous system, the epigenetic regulation of gene expression is critical for developmental processes and for complex brain functions. Dysregulation of the epigenetic profile of nerve cells can be detrimental, and has been associated with neurodevelopmental and neurodegenerative disorders. The dynamics and reversibility of epigenetic regulation may provide new options for potential diagnostic and for the development of epigenetic drug treatment.For finding the target proteins associated with brain disorders, this project plans to use ELISA to analyze specific target protein expression differences of endothelial cells (E), astrocytes(A) and neuron(N).Furthermore, the molecular biology and biochemical techniques such as RT-PCR, Western Blot, Immunofluorescence cytochemistry staining, and methylation specific PCR would be used to observe the influences of epigenetic regulation on the target proteins expression in vitro.Finally, high throughput DNA methylation chip will be used to detect clinical samples. Using Transwell coculture system, we has established a novel four-dimensional model in primary culture of tricellular elements, E, A, and N, from same-origin of brain or spinal cord. This four

英文关键词： neuroinflammation；multiple sclerosis；epigenetic regulation；DNA methylation；PGE2