Regulation of Mitochondrial Redox Systems in T-lymphocytes During Hypertension

描述（由申请人提供）：本项目将研究线粒体超氧化物(o2 ？-)对t淋巴细胞活化在高血压发病中的作用。活性氧，主要是o2 ？-，已被阐明为血管紧张素II （AngII）依赖性高血压的关键分子中间体。最近，免疫系统，特别是t淋巴细胞活化，已被证明是血管内皮介导的高血压的病因，它加剧了促炎环境；然而，o2的作用是什么？- in介导血管刺激t淋巴细胞内信号通路尚不清楚。在其他细胞类型中，线粒体已被确定为血管诱导的O2的关键来源。-产生，如线粒体定位的O2 ？清除酶锰超氧化物歧化酶（MnSOD）减弱血管i介导的O2？——通量。我们的初步数据表明，线粒体也是氧气的来源之一。-在血管刺激t淋巴细胞中。在此，我们假设线粒体产生的O2 ？-介导细胞内t淋巴细胞信号传导，导致其在血管内皮介导的高血压过程中激活和增强细胞因子的产生。在Specific Aim 1中，我们将研究线粒体O2的因果来源。-在AngII刺激小鼠原代t淋巴细胞中。有希望的候选来源包括NADPH氧化酶（NOX）酶、MnSOD和电子传递链。特异性目的2将研究AngII如何在t淋巴细胞中进行信号转导。利用微阵列和细胞因子阵列技术，我们将对信号转导途径进行全面分析，我们将获得特定的第二信使和转录因子来进一步验证。我们将使用mnsod缺失和mnsod过表达的小鼠模型来了解线粒体O2 ？-到血管i介导的t淋巴细胞胞内信号传导。最后，Specific Aim 3将检查线粒体O2 ？-在体内t淋巴细胞中通过检测线粒体O2 ?紊乱小鼠的平均动脉压（MAP）和心率（HR）。-特别是在t淋巴细胞中，我们将获得与高血压直接相关的体内终点。总的来说，该项目将进一步了解免疫系统对高血压的贡献，以及这种贡献背后的氧化还原机制，并为目前尚未认识到的高血压药物治疗提供潜在的新靶点。

DESCRIPTION (provided by applicant): This project will investigate the influence of mitochondrial superoxide (O 2?-) in T-lymphocytes on their activation in the pathogenesis of hypertension. Reactive oxygen species, mainly O 2 ?-, have been elucidated as key molecular intermediates in angiotensin II (AngII)-dependent hypertension. Recently, the immune system, particularly T-lymphocyte activation, has proven causal in AngII-mediated hypertension by exacerbating a pro- inflammatory environment; however, the role of O 2 ?- in mediating intracellular signaling pathways in AngII-stimulated T-lymphocytes remains unclear. In other cell types, mitochondria have been identified as a key source of AngII-induced O2 ?- production, as over-expression of the mitochondrially-localized O2 ?- scavenging enzyme manganese superoxide dismutase (MnSOD) attenuates AngII-mediated O2?- flux. Our preliminary data suggest mitochondria are also a source of O2 ?- in AngII-stimulated T-lymphocytes. Herein, we hypothesize that mitochondrially-produced O2 ?- mediates intracellular T-lymphocyte signaling leading to their activation and enhanced production of cytokines during AngII-mediated hypertension. In Specific Aim 1, we will investigate a causal source of mitochondrial O2?- in mouse primary T-lymphocytes stimulated with AngII. Promising candidate sources include the NADPH oxidase (NOX) enzymes, MnSOD, and the electron transport chain. Specific Aim 2 will examine how AngII carries out signal transduction in T-lymphocytes. Using microarray and cytokine array technology, we will perform a comprehensive analysis of signal transduction pathways, which we will garner specific second messengers and transcription factors to further validate. We will employ the use of MnSOD-deficient and MnSOD-overexpressing mouse models to understand the contribution of mitochondrial O2 ?- to AngII-mediated T-lymphocyte intracellular signaling. Finally, Specific Aim 3 will examine physiological parameters of altered mitochondrial O2 ?- in T-lymphocytes in vivo. By examining mean arterial pressure (MAP) and heart rate (HR) in mice with perturbed mitochondrial O2 ?- specifically in T-lymphocytes, we will obtain in vivo endpoints that will provide a direct connection to hypertension. Overall, this project will further the understanding of the immune system contribution to hypertension, the redox based mechanisms underlying this contribution, and provide potential new targets for pharmaceutical therapy that are currently unacknowledged in hypertension.