缓激肽及其B2受体活化调节内皮祖细胞衰老的作用及机制研究

项目名称： 缓激肽及其B2受体活化调节内皮祖细胞衰老的作用及机制研究

项目编号： No.81470401

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 姚玉宇

作者单位： 东南大学

项目金额： 73万元

中文摘要： 应激诱导细胞衰老（SIP）致内皮祖细胞（EPCs）数量及功能下降，与心血管病危险因素负相关。缓激肽（BK）B2受体（B2R）活化抑制内皮细胞SIP，其机制不明。我们预试验新发现BK/B2R活化抑制EPCs SIP，其机制与抑制视网膜母细胞瘤基因（RB）功能相关。申请者拟使用B2R激动剂、抑制剂、B2R腺病毒转染，siRNA干扰，RNA及蛋白芯片筛选等方法，研究BK/B2R活化抑制EPCs SIP。使用信号通路中多个调控单元的阻断剂和激动剂对相关的信号通路进行研究。使用糖尿病小鼠颈动脉内膜损伤模型，移植B2R高表达EPCs，活体成像，动态观察EPCs在损伤内膜的存活及衰老，阐明BK/B2R活化抗衰老通过B2R/PI-3K/Akt/周期蛋白/RB和/或B2R转激活EGFR/周期蛋白/RB途径。通过BK/B2R轴调节EPCs抗SIP，对改善EPCs功能, 治疗冠心病、糖尿病血管病变具有重要意义。

中文关键词： 内皮祖细胞；氧化应激；细胞衰老；缓激肽；B2受体

英文摘要： Stress-induced premature senescence (SIP) could impair endothelial progenitor cells (EPCs) function and proliferative capability, there is a negative correlation between EPCs function and cardiovascular risk factors. Bradykinin protects endothelial cells from SIP, however, the exact molecular mechanism of SIP and their role in EPCs dysfunction still needs to be elucidated. Our preliminary data showed BK inhibited H2O2-induced EPCs senescence through B2 receptor (B2R) activation, the level of RB1 transcripts was decreased by approximately one hundred and seventy-six fold by human cellular senescence PCR array. Here, we describe a novel mechanism of BK/B2R signal pathway activation leading to EPCs antisenescence. EPCs will be studied using Adenovirus transfection, RNA interference, real time PCR, Westernblotting, human cellular senescence PCR array, cell cycle control phospho antibody array and signal transduction inhibitors and agonists. In db/db diabetic mice, Ad.B2R modified EPCs will be loaded with near-infared dye and image in a wire-induced carotid injury model over a time course of 2 weeks using Maestro in vivo imaging system, to observe EPCs survival and senescence. We try to clarify the mechanism of BK/B2R activation EPCs inhibited SIP through B2R/ PI-3K-Akt /cyclinD/E or EGFR transactivation /cyclinD/E/RB pathway. Identification of the exact mechanisms of BK/B2 activation inhibition EPCs senescence may have important implications on the development of novel molecular therapies for the prevention of coronary heart disease and diabetic angiopathies

英文关键词： endothelial progenitor cell;oxidative stress;senescence;bradykinin;B2 reeptor