The anti-inflammatory response after acute kidney injury

描述（由申请人提供）： 急性肾损伤（阿基）发生在20%的住院患者中，并增加了死亡风险。阿基的死亡通常是由于全身性并发症。呼吸衰竭是阿基的一种特别有害的并发症，可使患者死亡率增加至60%至80%。阿基导致呼吸衰竭的原因尚不清楚，但可能是由于炎症。我们在小鼠中的数据表明，阿基是一种促炎状态，其特征在于2小时时血清促炎介质（细胞因子）增加，4小时时发生肺损伤。小鼠阿基后的肺损伤的特征在于肺中性粒细胞积聚和肺毛细血管渗漏。在这项研究中，我们提出阿基是一种促炎事件，通常通过产生IL-10启动抗炎反应。IL-10是一种有效的抗炎细胞因子，可抑制促炎介质的产生。我们认为，称为巨噬细胞的细胞是阿基后IL-10产生的关键来源。我们的总体假设是，巨噬细胞中IL-10的产生对于抑制阿基后促炎细胞因子的产生和肺损伤是必要的。提出了了解小鼠阿基后正常抗炎反应的研究，目的是确定促进炎症和肺损伤消退的疗法。 阿基后的伤害在具体目标1中，我们将确定阿基后脾脏中的巨噬细胞是否增加IL-10的产生。我们假设巨噬细胞是主要来源，IL-6是这种反应的关键介质。GFP报告小鼠（产生IL-10的细胞为GFP阳性）和流式细胞术也将用于确定脾巨噬细胞是否产生IL-10。将在体内和体外测试IL-6在介导脾IL-10产生中的作用。在具体目标2中，我们将确定IL-10是否是减少阿基中促炎细胞因子产生和限制肺损伤所必需的。我们假设IL-10限制了阿基后促炎细胞因子的产生和肺损伤。为了确定阿基后IL-10的有益作用，将在IL-10缺乏的阿基中确定血清细胞因子、器官细胞因子产生和肺损伤。将在野生型和IL-10缺陷小鼠中测试IL-10在阿基中的治疗潜力;将在损伤前或损伤后的某个时间施用IL-10以确定IL-10是否可 这是一种潜在的治疗方法。在具体目标3中，我们将确定产生IL-10的巨噬细胞是否促进阿基介导的肺损伤的消退。我们假设产生IL-10的巨噬细胞限制阿基后的肺损伤。将在巨噬细胞中缺失IL-10的小鼠中评估产生IL-10的巨噬细胞在阿基中的作用;还将进行产生IL-10或缺乏IL-10的巨噬细胞的给药。

DESCRIPTION (provided by applicant): Acute kidney injury (AKI) occurs in 20% of hospitalized patients and increases the risk of death. Death from AKI is typically due to systemic complications. Respiratory failure is an especially detrimental complication of AKI and can increase patient mortality to 60 to 80%. The reason AKI causes respiratory failure is unclear, but may be due to inflammation. Our data in mice demonstrate that AKI is a pro-inflammatory state that is characterized by increased serum proinflammatory mediators (cytokines) at 2 hours and lung injury that occurs by 4 hours. Lung injury after AKI in mice is characterized by lung neutrophil accumulation and lung capillary leak. In this grant, we propose that AKI is a proinflammatory event that normally initiates a counter inflammatory response via production of IL-10. IL-10 is a potent anti-inflammatory cytokine that inhibits production of proinflammatory mediators. We suggest that cells known as macrophages are the key source of IL-10 production after AKI. Our overall hypothesis is that IL-10 production in macrophages is necessary to contain proinflammatory cytokine production and lung injury after AKI. Studies to understand the normal counter inflammatory response after AKI in mice are proposed with the goal of identifying therapies that facilitate resolution of inflammation and lung injury after AKI. In Specific Aim 1, we will determine whether macrophages in the spleen increase IL-10 production after AKI. We hypothesize that macrophages are the major source and that IL-6 is a key mediator of this response. GFP reporter mice (IL-10 producing cells are GFP positive) and flow cytometry will also be used to determine if splenic macrophages produce IL-10. The role of IL-6 in mediating splenic IL-10 production will be tested in vivo and in vitro. n Specific Aim 2, we will determine if IL-10 is necessary to reduce proinflammatory cytokine production and limit lung injury in AKI. We hypothesize that IL-10 limits proinflammatory cytokine production and lung injury after AKI. To determine the beneficial role of IL-10 after AKI serum cytokines, organ cytokine production, and lung injury will be determined in AKI with IL-10 deficiency. The therapeutic potential of IL-10 in AKI will be tested in wild type and IL-10 deficiet mice; IL-10 will be administered prior to injury or at times after injury to determine if IL-10 may be a potential treatment. In Specific Aim 3, we will determine if IL-10 producing macrophages facilitate resolution of AKI- mediated lung injury. We hypothesize that IL-10 producing macrophages limit lung injury after AKI. The role of IL-10 producing macrophages in AKI will be assessed in mice with deletion of IL-10 in macrophages; administration of IL-10 producing or deficient macrophages will also be performed.