The Role of Caspases in Warm and Cold Ischemia

Caspases 在热缺血和冷缺血中的作用

• 批准号: 7474767
• 负责人: Alkesh Harihar Jani
• 金额: $ 13.11万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7474767
• 关键词: Acute; Acute Kidney Failure; Allografting; Antigens; Apoptosis; Apoptotic; Applications Grants; Brush Border; Caspase; Caspase Inhibitor; Caspase-1; Cell Death; Cells; Cessation of life; Chronic; Chronic rejection of renal transplant; Cryopreservation; Data; Development; Epithelial Cells; Functional disorder; Gene Expression; Grant; Immune Sera; Immunohistochemistry; Incidence; Inflammatory; Injury; Interferon Type II; Interferons; Interleukin-18; Ischemia; Kidney; Kidney Diseases; Kidney Transplantation; MHC Class I Genes; Mediating; Mediator of activation protein; Messenger RNA; Mitochondria; Modeling; Morphology; Mus; Necrosis; Organ; Organ Preservation; Organ Preservation Solutions; Perfusion; Physiological reperfusion; Production; Proteins; Protocols documentation; Publishing; Reperfusion Therapy; Research Personnel; Role; Transplantation; Tubular formation; Warm Ischemia; base; caspase-3; caspase-8; caspase-9; clinically relevant; cytochrome c; cytokine; delayed graft function; immunogenicity; implantation; inhibitor/antagonist; interferon-alpha B; interstitial; mRNA Expression; prevent; programs; receptor; research study; tubular necrosis

Delayed graft function (DGF) independently predicts reduced 1- and 5- year kidney transplant survival and contributes to the development of chronic allograft nephropathy (CAN). DGF is caused by both cold and warm ischemia. Cold ischemia occurs during both pulsatile machine perfusion and static cold storage of kidneys. This grant will focus on the pathophysiology of cold and warm ischemia and specifically the role of caspases. Caspase-3 is a major mediator of apoptotic and necrotic cell death. Caspase-1 converts the pro- inflammatory cytokines IL-1beta and IL-18 to their active forms. The overall hypothesis is that caspases mediate proximal tubular damage in both cold and warm ischemia. In a model of cold ischemia the contribution of caspase-3 to proximal tubular apoptosis as well as severe brush border injury will be assessed. Gene expression, protein and activity of caspases-3, 7, 9 (mitochondrial pathway of apoptosis) and 8 (death receptor pathway of apoptosis) will be determined during both static and pulsatile cold ischemia. Immunohistochemistry for activated caspase-3 in proximal tubules will be performed. The effect of caspase inhibition on kidney morphology will be assessed. The findings of these experiments will contribute to the understanding of cold ischemia injury and organ preservation. Caspase-1-mediated production of IL-18 mediates ischemic ARF. IL-18 is a potent inducer of interferbn gamma, which is a powerful inducer of MHC expression. Caspase-1 and IL-18-mediated increase in interferon gamma expression in the kidney and MHC expression on proximal tubules will be determined in a model of warm ischemia. Interferon gamma and MHC class I and II mRNA expression in the kidney will be assessed. The effect of caspase-1, IL-18 and interferon gamma inhibition on MHC expression will be evaluated. Reduced MHC expression is associated with decreased kidney graft immunogenicity and has important implications for the deleterious effect of DGF on both short and long-term allograft function.

延迟的移植功能（DGF）独立预测1年和5年肾脏移植生存的降低和 有助于慢性同种异体移植肾病（CAN）的发展。 DGF是由冷和 温暖的缺血。冷缺血在脉冲机灌注和静态冷藏期间发生 肾脏。该赠款将重点放在冷和温暖缺血的病理生理上，尤其是 caspase。 caspase-3是凋亡和坏死细胞死亡的主要介体。 caspase-1转换支持 炎症细胞因子IL-1Beta和IL-18的活性形式。总体假设是caspase 介导冷和温暖缺血的近端管状损伤。在冷缺血模型中 caspase-3对近端管状细胞凋亡以及严重的刷边界损伤的贡献将是 评估。 caspases-3，7，9的基因表达，蛋白质和活性（凋亡的线粒体途径） 和8（凋亡的死亡受体途径）将在静态和脉冲冷时确定 缺血。将对近端小管中激活的caspase-3进行免疫组织化学。效果 将评估caspase对肾脏形态学的抑制作用。这些实验的发现将 有助于理解冷缺血损伤和器官保存。 caspase-1介导的 IL-18介导的产生缺血性ARF。 IL-18是干扰伽玛的有效诱导剂，这是 MHC表达的强大诱导剂。 caspase-1和IL-18介导的干扰素伽玛的增加 肾脏和MHC在近端小管上的表达将在温暖的模型中确定 缺血。将评估肾脏中的干扰素γ和MHC I类和II类mRNA表达。这 将评估caspase-1，IL-18和干扰素γ对MHC表达的影响。减少 MHC表达与肾脏移植免疫原性降低有关，并具有重要意义 DGF对短期和长期同种异体移植功能的有害影响。