甘青铁线莲中新的活性成分APG激活GSK-3β/Nrf2通路减轻脑缺血/再灌注损伤的机制研究

项目名称： 甘青铁线莲中新的活性成分APG激活GSK-3β/Nrf2通路减轻脑缺血/再灌注损伤的机制研究

项目编号： No.81503285

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 张伟

作者单位： 中国人民解放军第四军医大学

项目金额： 18万元

中文摘要： 缺血性脑卒中是严重危害人类健康的重大疾病。目前临床尚无理想的治疗药物或措施。寻找新型高效安全、作用机制清晰的脑卒中防治药物是治疗该疾病的迫切需求。申请者前期从传统藏药甘青铁线莲中获得一种黄酮苷类活性成分APG，首次发现APG能够显著降低大鼠脑中动脉栓塞(MCAO)造成的缺血再灌注损伤，减少脑梗死面积，改善神经行为学评分。预实验研究发现APG干预能够通过增加脑缺血损伤后GSK-3β磷酸化，降低内源性ROS水平，增加T-SOD的活性，抑制细胞凋亡。提示APG可能通过激活GSK-3β/Nrf2通路，调节缺血/再灌注损伤介导的氧化应激，进而发挥脑保护作用。因此，本项目拟在前期工作的基础上，采用基因转染技术，基因沉默技术，流式细胞术、免疫组化、免疫电镜技术等方法，从整体、细胞、分子水平阐明APG的脑保护机制，为其治疗缺血性脑卒中的临床应用提供理论依据。

中文关键词： 甘青铁线莲；脑缺血/再灌注损伤；糖原合成酶激酶-3β；NF-E2；相关因子2；；作用机制

英文摘要： Ischemic stroke is a major disease which has seriously jeopardized human health.At present, there are no ideal clinical treatments or measures. The process of discovering novel efficient and safe drugs was more urgent. The applicant obtained the active ingredient named APG from the Tibet herbal medicine Clematis tangutica. And found for the first time that APG has significantly reduced cerebral ischemia/reperfusion injury, reduced infarct size and improved neurological behavior score. The result of pre-experiment showed that APG pretreatment increased GSK-3β phosphorylation after reperfusion on ischemic injury, decreased endogenous ROS levels, increased T-SOD activities, and inhibited cell apoptosis. This suggests that APG may activated GSK-3β/Nrf2 pathway, accommodated ischemia reperfusion-mediated oxidative stress, exerted a protection on the ischemic brain. Thus, we planned to apply the Gene Transfection, Gene Silencing, Flow Cytometer and Immunohistochemisty method and technic to explore the potential role of GSK-3β/Nrf2 pathway in brain protection of APG on the animal, cellular and molecular levels. This project provide theoretical basis for APG treating ischemic stroke in clinical in the future

英文关键词： Clematis tangutica;cerebral ischemia/reperfusion injury;GSK-3β;Nrf2;Mechanism