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AT 1 R和 AT 2 R导致动物模型中炎症的诱导。衰老对血管紧张素Ⅱ 1型受体表达的影响 人类的AT 2 R以及AT 1 R和AT 2 R的变化对老年人炎症增加的贡献， 以前没有研究过。我们的初步证据表明，虚弱的老年人有上调， AT 1 R表达下调，AT 2 R表达下调，IL 6参与了这种失衡。 我们推测，人类衰老与AT 1 R的上调和AT 2 R的下调有关 在免疫系统细胞中的表达和功能。我们假设这些受体的变化有助于 免疫系统细胞吞噬能力降低和炎性细胞因子产生增加 这将进一步增加AT 1 R和AT 2 R表达的差异。 为了验证这些假设，我们提出了一个全面的研究AT 1 R和AT 2 R使用免疫 系统细胞（淋巴细胞和单核细胞）从年轻，健康成人（年龄20-30）和四个比较 包括（1）健康的老年人（年龄70-90岁），（2）健康的老年人（年龄70-90岁）， AT 1 R阻滞剂，（3）虚弱的老年人（70-90岁），（4）接受AT 1 R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，用于以下建议的研究。 研究：1.测量AT 1 R和AT 2 R的基因表达、蛋白质合成和信号通路的变化 在来自年轻对照组（20- 30岁）和四个对照组（每组N=33）的免疫系统细胞中 使用Q-PCR、蛋白质印迹、共聚焦显微镜、流式细胞术和Bio-Phosphoprotein Cellular 信号传导测定。 2.血管紧张素受体阻断剂对单核细胞吞噬功能的影响及其与年龄的关系 通过将来自相同个体的免疫系统细胞与特异性AT 1 R和/或AT 2 R阻断剂一起孵育来发挥作用 并在基线和基线时用吞噬测定测量单核细胞吞噬功能的变化。 对治疗的反应。 3.通过孵育免疫细胞，评估AT 1 R和AT 2 R对老年人细胞因子产生的贡献。 系统细胞与特定的AT 1 R和/或AT 2 R阻断剂和测量细胞因子， 基线时和治疗后的ELISA和Bio-ELISA细胞因子测定。 4.通过孵育免疫反应来评估炎症对AT 1 R和AT 2 R表达和功能的反馈。 来自相同受试者的系统细胞与IL-6。将使用Q-PCR和蛋白质印迹来定量细胞中的变化。 AT 1 R和AT 2 R的表达响应于IL-6处理。

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.