FIBRONECTION IN HEPATIC ISCHEMIA REPERFUSION INJURY

肝缺血再灌注损伤中的纤维连接

• 批准号: 6887678
• 负责人: Ana J. Coito
• 金额: $ 34.76万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6887678
• 关键词: biological signal transduction; cell adhesion; cell migration; cryopreservation; cytokine; enzyme activity; fibronectins; free radical oxygen; gene expression; genetically modified animals; inflammation; integrins; laboratory mouse; laboratory rat; leukocyte activation /transformation; leukocytes; liver function; liver ischemia /hypoxia; liver transplantation; metalloendopeptidases; nitric oxide synthase; oxidative stress; protein structure function; reperfusion; tumor necrosis factor alpha

DESCRIPTION (provided by applicant): lschemia reperfusion (I/R) injury remains a major limitation in orthotopic liver transplantation (OLT), particularly with marginal grafts. Very little is known about the mechanisms involved on leukocyte recruitment into sites of inflammatory stimulation in liver. Based upon evidence from our previous studies, we hypothesize that fibronectin (FN) is an integral part of the antigen independent cascade leading to liver I/R injury, likely providing a spatial address at which appropriate co-stimulatory signals can initiate leukocyte signaling and recruitment. We expect this proposal will unveil novel mechanisms and potential targets against I/R injury. Therefore, we propose to address the following specific aims: 1 . To analyze the function of distinct FN-alpha4beta1/alpha5beta1interactions upon leukocyte recruitment and cytokine release in liver I/R injury: Livers from genetically fat Zucker and non-steatotic SD rats will be stored at 4C in UW before being transplanted into syngeneic lean Zucker/SD rats, and the three distinct FN (EIIIA)-alpha4beta1, FN (CS1)-alpha4beta1,and FN (RGD)-alpha5beta1 interactions will be targeted with specific peptides and function-blocking mAbs. The distinct therapeutic manipulation upon (i) hepatic function and survival; (ii) leukocyte adhesion/migration (in vivo and in-vitro), and (iii) cytokine expression, will be probed. 2. To dissect mechanisms of FN-alpha4beta1/alpha5beta1 integrin action upon metalloproteinase-2 (MMP-2) and metalloproteinase-9 (MMP-9) expression in liver I/R Injury: We plan to (i) determine the contribution of distinct FN cell binding sites upon MMP-2 and MMP-9 expression/activation, and respective inhibitors (TIMP-2 and TIMP-1) in rat OLT; (ii) identify the individual sources of MMP-2 and MMP-9 in liver grafts; and (iii) evaluate the functional significance of MMP-2 and MMP-9 expression in the mechanisms of FN-integrin interactions upon liver I/R injury. 3. To explore the interplay between inducible nitric oxide synthase (iNOS), MMPs, and cytokines in the context of FN in liver I/R injury: First, the functional significance of iNOS expression will be probed in vivo with 1400W, a specific iNOS inhibitor. Second, we will dissect the interplay between TNF-alpha or other relevant cytokines, and iNOS upon FN regulation of MMP expression/activation. Third, we will unveil intracellular mechanisms, such as MAPKinase transduction pathways upon leukocyte adhesion to fibronectin and MMP expression.

描述（由申请人提供）： LSCHEMIA再灌注（I/R）损伤仍然是原位肝移植（OLT）的主要局限性，尤其是边缘移植物。关于白细胞募集到肝脏炎症部位所涉及的机制知之甚少。根据我们先前研究的证据，我们假设纤连蛋白（FN）是导致肝I/R损伤的抗原独立级联反应的组成部分，可能会提供空间地址，适当的共同刺激信号可以启动白细胞信号和招募。我们预计该建议将推出新的机制和针对I/R伤害的潜在目标。因此，我们建议解决以下特定目标：1 。分析肝脏I/R损伤中白细胞募集和细胞因子释放时不同的FN-Alpha4Beta1/alpha5beta1间隔：遗传性脂肪Zucker的肝脏和非steateatotic SD大鼠的肝脏将在UW中以4C储存在UW中，然后将其移植到Syngence flats中，并将三分之一的SD rats和SD rats和Sd rats sd rats rats rats。 （EIIIA）-alpha4beta1，fn（CS1）-Alpha4beta1和Fn（RGD）-Alpha5beta1相互作用将以特定的肽和功能阻滞的mAB靶向。 （i）肝功能和生存的独特治疗操作； （ii）将探测白细胞粘附/迁移（体内和体内）和（iii）细胞因子的表达。 2。剖析肝I/r损伤中Fn-Alpha4Beta1/alpha5beta1整合素对金属蛋白酶-2（MMP-2）和金属蛋白酶9（MMP-9）表达的机制的作用：我们计划（i）确定（i）确定（i）确定（i）对MMP-2和MMP-2的贡献（i）确定（i） timp-1）在大鼠olt中； （ii）确定肝移植中MMP-2和MMP-9的各个来源； （iii）评估MMP-2和MMP-9表达在肝I/R损伤时FN-整合素相互作用机理中的功能意义。 3。探索在肝I/R损伤中FN的背景下诱导型一氧化氮合酶（INOS），MMP和细胞因子之间的相互作用：首先，将用1400W的iNOS表达的功能意义在1400W中探测为特定的Inos抑制剂。其次，我们将剖析TNF-α或其他相关细胞因子之间的相互作用，以及在FN调控MMP表达/激活后的INOS。第三，我们将揭露细胞内机制，例如白细胞粘附到纤连蛋白和MMP表达时的Mapinase转导途径。