Nitric oxide and mitochondrial biogenesis in sepsis

这是一个修正的应用程序，研究的基本机制，一氧化氮（NO）的作用，在线粒体 败血症时肝脏的生物合成NO合酶的诱导是脓毒症诱导的免疫应答的基础 但活性氮和氧的产生（RNS，ROS）是一种应激， 在多器官功能衰竭（MOF）的发病机制中是重要的。我们发现 实验性脓毒症中的RNS和ROS产生损伤线粒体DNA（mtDNA），从而损害 线粒体转录、蛋白质组保真度和线粒体功能。这种线粒体损伤 刺激细胞补偿，涉及神经生物发生，这需要激活线粒体 转录因子A（mtTFA）和两个核转录因子，核呼吸因子-1和-2 （NRF-1和NRF-2）和共活化剂PGC-1。这些独特的细胞反应，在核控制下， RNS和ROS调节线粒体生物学的关键方面，也是促生存的一个方面。虽然我们 我们的数据显示，对于炎症中生物发生的调节或脓毒症中生物发生的破坏， 很明显，线粒体病理学不仅仅局限于损伤线粒体的NO化学， 杀死细胞。因此，我们建议测试iNOS刺激线粒体生物合成的假设， 对抗脓毒症中的细胞坏死，因此，生物发生的调节是脓毒症的重要决定因素。 细胞存活为了验证这一假设，我们提出了三个具体目标：目标1：衡量 诱导型一氧化氮合酶在脓毒症肝线粒体DNA和蛋白质损伤中的作用 目的2：明确iNOS在线粒体转录因子A中的重要性 （Tfam）激活和恢复肝线粒体DNA拷贝数和转录在野生型和 脓毒症中iNOS基因敲除小鼠;目的3：确定iNOS对核转录的贡献 在野生型和iNOS敲除小鼠中通过NRF-1和NRF-2表达激活脓毒症中的生物发生。 这项工作将提供一个更好的机械理解的核线粒体通信过程中， 宿主的炎症反应，这将有助于促进新的分子策略，以评估的能力， 线粒体反应标记物来预测MOF并指导干预措施以预防和最终治疗 脓毒症引起的MOF。

This is an amended application to study the basic mechanisms of action of nitric oxide (NO) in mitochondrial biogenesis in the liver in sepsis. NO synthase induction is fundamental to the sepsis-induced immune response but the production reactive nitrogen and oxygen species (RNS, ROS) is a stress that drives mitochondrial dysfunction and is important in the pathogenesis of multiple organ failure (MOF). We have discovered that RNS and ROS production in experimental sepsis damage mitochondrial DNA (mtDNA), thereby impairing mitochondrial transcription, proteome fidelity, and mitochondrial function. This mitochondrial damage stimulates cellular compensation involvingmitochondrialbiogenesis, which requires activation ofmitochondrial transcription factor A (mtTFA) and two nuclear transcription factors, nuclear respiratory factor-1 and -2 (NRF-1 and -2),and a co-activator, PGC-1. These unique cell responses, under nuclear control, signify a crucial side of mitochondrial biology modulated by RNS and ROS, and one that is pro-survival. Though we still know very little about regulation of biogenesis in inflammation or its disruption in sepsis, our data show clearly that mitochondrial pathology is not limited simply to the NO-chemistry that damages mitochondria and kills cells. Therefore, we propose to test the hypothesis that iNOS-stimulated mitochondrial biogenesis opposes cell necrosis in sepsis, and thereby, regulation of biogenesis is an important determinant of cell survival. To test this hypothesis we propose three Specific Aims: Aim 1: Measure the contribution of iNOS to the pathogenesis of damage to hepatic mitochondrial DNA and proteins in sepsis using wild type and iNOS knockout mice; Aim 2: Define the importance of iNOS in mitochondrial transcription factor A (Tfam) activation and the restoration of hepatic mtDNA copy number and transcription in wild type and iNOS knockout mice in sepsis; Aim 3: Determine the contribution of iNOS to nuclear transcriptional activation of biogenesis in sepsis via NRF-1 and NRF-2 expression in wild type and iNOS knockout mice. This work will provide a better mechanistic understanding of the nuclear-mitochondrial communication during the host inflammatory response, which should help foster new molecular strategies to assess the ability of mitochondrial response markers to predict MOF and to guide interventions to prevent and eventually to treat sepsis-induced MOF.