Preclinical evaluation of resveratrol in sepsis-induced renal injury

白藜芦醇治疗脓毒症肾损伤的临床前评价

• 批准号: 8064784
• 负责人: Joseph Hunter Holthoff
• 金额: $ 2.97万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8064784
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Antioxidants; Blood capillaries; Chemicals; Clinical; Creatinine; Data; Development; Diagnosis; Dose; Epithelial; Epithelial Cells; Epithelium; Event; Functional disorder; Generations; Hour; Injury; Kidney; Knowledge; Laboratories; Ligation; Link; Mediator of activation protein; Modeling; Morphology; Mus; New Agents; Oxidants; Oxygen; Patients; Perfusion; Peroxonitrite; Puncture procedure; Reaction; Reactive Nitrogen Species; Reactive Oxygen Species; Renal function; Renal tubule structure; Reporting; Research Proposals; Resveratrol; Role; Scientist; Sepsis; Serum; Stream; Stress; Superoxide Dismutase; Therapeutic; Time; Training; Tubular formation; United States; United States National Institutes of Health; Video Microscopy; base; capillary; cell injury; cytokine; density; ebselen; in vitro testing; innovation; intravital video microscopy; mortality; new therapeutic target; novel therapeutic intervention; overexpression; polyphenol; preclinical evaluation; prevent; response; septic

DESCRIPTION (provided by applicant): I propose that during sepsis decreased peritubular capillary perfusion leads to a pro-oxidant peritubular microenvironment, which favors the generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS). My preliminary data show that acute treatment with the naturally occurring anti-oxidant polyphenol, resveratrol, prevented the generation of tubular RNS and the decline in peritubular capillary perfusion observed at 18h following induction of sepsis using the cecal ligation and puncture (CLP) in a murine model of sepsis. Moreover, I have generated direct evidence that resveratrol can scavenge RNS. Based on these findings, I hypothesize that the RNS scavenging ability of resveratrol will protect the mouse renal tubules from the pro-oxidant peritubular microenvironment produced during CLP-induced. Aim 1 will determine the efficacy of resveratrol to protect renal tubular epithelia from RNS using primary cultures of mouse renal tubular epithelial cells. Aim 2 will examine the therapeutic potential of resveratrol to prevent tubular epithelial cell injury caused by CLP. Dose-response curves for resveratrol over the time course leading to renal injury will be generated and analyzed for the effects on serum cytokine profile, functional peritubular capillary density, volumetric flow, tubular oxidant generation (each quantified simultaneously using intravital videomicroscopy), and renal injury (morphology and serum BUN/creatinine). Aim 3 will establish RNS as the key mediator of CLP-induced renal injury in the mouse using iNOS mice and in mice that over-express superoxide dismutase (S0D1) to reduce the levels of NO and O2" available for RNS (ONOO") generation, respectively. A very innovative aspect of this study is the use of intravital video microscopy (IWM) to study in real time the mechanisms by which capillary dysfunction develops and leads to tubular injury following CLP. This research proposal will contribute significant to my overall training to become a translational clinical scientist. Each year and more than 210,000 people die from sepsis-related complications in the United States. Acute kidney injury occurs in 20%-50% of septic patients and the mortality of these septic patients is extremely high (~75%) because current therapy is mostly supportive and largely ineffective. The major obstacle to the development of new therapeutic approaches for sepsis-induced kidney injury is the lack of knowledge regarding critical events that can be targeted after the initiation of sepsis. This proposal investigates new therapeutic targets to treat sepsis and has the potential to identify new agents to reduce mortality of this devastating condition.

描述（由申请人提供）：我提出，在脓毒症期间，减少的管周毛细血管灌注导致促氧化剂管周微环境，其有利于活性氧物质（ROS）和活性氮物质（RNS）的产生。我的初步数据表明，急性治疗与天然存在的抗氧化剂多酚，白藜芦醇，防止产生的肾小管RNS和肾小管周围毛细血管灌注下降观察到在18小时后，诱导败血症使用盲肠结扎和穿孔（CLP）在小鼠模型中的败血症。此外，我已经产生了直接的证据表明，白藜芦醇可以抑制RNS。基于这些发现，我推测白藜芦醇的RNS清除能力将保护小鼠肾小管免受CLP诱导过程中产生的促氧化剂管周微环境的影响。目的1利用原代培养的小鼠肾小管上皮细胞，观察白藜芦醇对RNS损伤肾小管上皮细胞的保护作用。目的2：研究白藜芦醇预防CLP引起的肾小管上皮细胞损伤的治疗潜力。将生成白藜芦醇在导致肾损伤的时间过程中的剂量-反应曲线，并分析其对血清细胞因子谱、功能性管周毛细血管密度、体积流量、肾小管氧化剂生成（使用活体视频显微镜同时定量）和肾损伤（形态学和血清BUN/肌酐）的影响。目的3将在使用iNOS小鼠和过表达超氧化物歧化酶（SOD 1）的小鼠中建立RNS作为CLP诱导的肾损伤的关键介质，以分别降低可用于RNS（ONOO）产生的NO和O2的水平。这项研究的一个非常创新的方面是使用活体视频显微镜（IWM）研究在真实的时间的机制，毛细血管功能障碍的发展，并导致肾小管损伤后CLP。这项研究计划将有助于我成为一名转化临床科学家的整体培训。 在美国，每年有超过21万人死于败血症相关并发症。急性肾损伤发生在20%-50%的脓毒症患者中，这些脓毒症患者的死亡率极高（约75%），因为目前的治疗大多是支持性的，而且大多无效。开发脓毒症诱导的肾损伤的新治疗方法的主要障碍是缺乏关于脓毒症开始后可以靶向的关键事件的知识。该提案研究了治疗脓毒症的新治疗靶点，并有可能确定新的药物来降低这种毁灭性疾病的死亡率。