AGE RELATED CHANGE IN ANGIOTENSIN RECEPTORS AND ITS ROLE IN CHRONIC INFLAMMATION

项目摘要： 衰老与炎症增强有关。血管紧张素受体AT1R和AT1R之间的比率改变 AT2R可诱导动物模型的炎症反应。衰老对血管紧张素Ⅱ1型受体和血管紧张素Ⅱ受体表达的影响 人类的AT2R以及AT1R和AT2R的变化对老年人炎症加剧的贡献 以前没有被研究过。我们的初步证据表明，虚弱的老年人有上调的 AT1R、AT2R表达下调和IL6参与了这种失衡。 我们假设人类虚弱的衰老与AT1R上调和AT2R下调有关 免疫系统细胞的表达和功能。我们假设这些受体的变化有助于 免疫系统细胞吞噬能力下降和炎性细胞因子产生增加 这将进一步加剧AT1R和AT2R表达的差异。 为了验证这些假说，我们提出了用免疫系统对AT1R和AT2R进行综合研究 青年健康成人(20-30岁)的系统细胞(淋巴细胞和单核细胞)及四种比较 由(1)健壮的老年人(70-90岁)，(2)健壮的老年人(70-90岁)组成的小组 AT1R阻滞剂，(3)虚弱的老年人(70-90岁)，(4)使用AT1R阻滞剂治疗的虚弱的老年人(70-90岁)。 我们将从这些受试者中收集淋巴细胞和单核细胞，用于以下建议 研究：1.检测AT1R和AT2R基因表达、蛋白质合成和信号通路的变化 青年对照组(20-30岁)和四个对照组(每组33个)的免疫系统细胞 应用Q-PCR、免疫印迹、共聚焦显微镜、流式细胞仪和Bio-Plex磷酸蛋白细胞 信号分析。 2.评价血管紧张素受体阻滞剂对单核细胞吞噬功能的年龄相关性差异及贡献 通过用特定的AT1R和/或AT2R阻断剂孵育来自相同个体的免疫系统细胞来发挥作用 并用吞噬试验测定基线和入院时单核细胞吞噬功能的变化。 治疗反应(S)。 3.通过孵育免疫来评估AT1R和AT2R对老年人细胞因子产生的贡献 使用特定AT1R和/或AT2R阻滞剂的相同个体的系统细胞，并用 在基线和治疗反应时进行ELISA法和Bio-Plex细胞因子分析(S)。 4.通过免疫孵育评价炎症对AT1R、AT2R表达和功能的反馈作用 同一受试者的系统细胞中含有IL-6。Q-pr和蛋白质印迹将被用来量化变化。 血管紧张素Ⅱ受体AT1R和AT2R在IL-6作用下的表达

Project Abstract: Aging is associated with enhanced inflammation. An altered ratio between angiotensin receptors AT1R and AT2R results in induction of inflammation in animal models. The effects of aging on the expression of AT1R and AT2R in humans and the contribution of changes in AT1R and AT2R to increased inflammation in the older have not been previously studied. Our preliminary evidence suggests that frail older adults have up-regulation of AT1R, down-regulation of AT2R expression and implicate for IL6 in this imbalance. We hypothesize that human frail aging is associated with up-regulation of AT1R and down-regulation of AT2R expression and function in immune system cells. We hypothesize that these receptor changes contribute to decreased immune system cells phagocytic capacity and to the increased production of inflammatory cytokines in older individuals which will further heighten the divergence in AT1R and AT2R expression. In order to test these hypotheses, we propose a comprehensive study of AT1R and AT2R using immune system cells (lymphocytes and monocytes) from young, healthy adults (age 20-30) and four comparison groups that consist of (1) robust, older adults (age 70-90), (2) robust, older adults (age 70-90) treated with AT1R blockers, (3) frail, older adults (age 70-90), (4) frail, older adults (age 70-90) treated with AT1R blockers. From these subjects we will collect lymphocytes and monocytes that will be utilized for the following proposed studies: 1. Measure changes in gene expression, protein synthesis, and signaling pathways of AT1R and AT2R in immune system cells from young control (20-30Y) and the four comparison groups (N=33 in each group) using Q-PCR, western blot, confocal microscopy, flow cytometry and Bio-Plex Phosphoprotein Cellular Signaling Assays. 2. Evaluate age-related difference and contribution of Angiotensin receptors blockade to monocytes phagocytic function by incubating immune system cells from the same individuals with specific AT1R and/or AT2R blockers and measuring changes in monocytes phagocytic function with Phagocytosis Assay at baseline and in response to treatment(s). 3. Evaluate contribution of AT1R and AT2R to cytokine production in the older individuals by incubating immune system cells from the same individuals with specific AT1R and/or AT2R blockers and measuring cytokines with ELISA and Bio-Plex cytokine assays at baseline and in response to treatment(s). 4. Assess the feedback of inflammation on AT1R and AT2R expression and function by incubating immune system cells from the same subjects with IL-6. Q-PCR and western blot will be used to quantify the change in expression of AT1R and AT2R in response to IL-6 treatment.