以端粒为靶点的抗氧化药物筛选和机制研究

Oxidative stress could change inflammatory signaling network to stimulate systemic inflammatory response and multiple organ failure. We found a new mechanism of oxidative damage that ROS induced telomere damage might cause inflammatory transcriptional reprogramming. Our previous experiments have shown that:(1)an original designed cell model that combined with telomeric heterochromatin properties and report gene can be used for high-throughput screening of drugs protects telomere function; (2)we used this model and found anti-oxidative drugs with telomere protection. We will further study:(1)whether the antioxidant effect of the drug is based on its role in protecting telomere structure and heterochromatic function; (2)how the drug protects against oxidative damage at telomeres, whether it is through the maintenance of telomeric heterochromatin in the dynamic regulation of transcriptional reprogramming to avoid ROS-induced inflammatory factors and cytokines over-expression; (3) we will verify the antioxidant drug in protecting telomere function in vivo via the zebrafish model of oxidative damage and telomere damage. We will use the following assay:southern-bolt、metaphase-PNA-FISH、immunofluorescence, Micro-array with RT-PCR methods. This project will contribute to the new mechanism of oxidative damage in molecular level and provide new idea for the treatment of critical ill patients. The telomere based screening model will favour to the explore of new antioxidant drugs and promote the secondary development of clinical drug.

传统观念认为氧化应激改变炎症信号网络，激发全身炎症反应和脏器衰竭。我们发现ROS介导的端粒损伤引发炎性转录重编程可能是氧化损伤的新机制。前期工作: 1) 原创性设计了以端粒异染色质属性结合报告基因的细胞模型；2）用该模型高通量筛选出保护端粒功能的抗氧化药物。本项目拟进一步证明：1）该药的抗氧化作用是通过保护端粒结构和异染色质功能实现的；2）上述药物保护端粒对抗氧化损伤的主要机制为：维护端粒的异染色质特性在转录重编程的动态调控，避免ROS介导炎症介质和细胞因子过度表达；3）在氧化损伤和端粒损伤的斑马鱼模型体内验证药物的抗氧化与保护端粒功能有关。上述研究将运用SOUTHERN-BLOT、有丝分裂中期染色体端粒功能、免疫荧光，Micro-array结合RT-PCR等方法。该项目将在分子水平阐明氧化损伤的新机制，为临床危重病治疗提供新思路。该端粒药筛模型有助于探索抗氧化新药，促进临床药物二次开发。