Matrix Metalloproteinases in Hepatic Ischemia and Reperfusion Injury

肝脏缺血和再灌注损伤中的基质金属蛋白酶

• 批准号: 8040076
• 负责人: Ana J. Coito
• 金额: $ 2.83万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 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 2010-05-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8040076
• 关键词: Acute; Address; Affect; Animals; Antibodies; Apoptosis; Basement membrane; Binding; Biological Preservation; Biological Process; Bone Marrow; Cells; Chronic; Clinical; Data; Development; Event; Extracellular Matrix; Failure; Free Radicals; Functional disorder; Funding; Gelatinase A; Gelatinase B; Gelatinases; Gene Transfer; Grant; Hepatic; Human; In Vitro; Incidence; Individual; Infiltration; Inflammatory; Injury; Lead; Leukocyte Trafficking; Leukocytes; Link; Liver; Liver Regeneration; Liver diseases; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Modeling; Mus; Natural regeneration; Obesity; Operative Surgical Procedures; Organ Donor; Outcome; Patients; Pattern; Peptide Hydrolases; Peptides; Physiological; Pilot Projects; Play; Population; Predisposition; Prevalence; Process; Property; Proteolysis; Public Health; Rattus; Regulation; Relative (related person); Reperfusion Injury; Risk; Role; Shock; Signal Transduction; Source; Staging; System; Testing; Therapeutic; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinases; Transplantation; Trauma; Vascular blood supply; Work; base; chemokine; cytokine; high risk; inhibitor/antagonist; insight; liver function; liver ischemia; liver transplantation; migration; mouse model; neutralizing antibody; novel therapeutics; overexpression; tumor

Hepatic ischemia reperfusion injury (IRI) occurs in all transplanted livers, in trauma, shock, and in elective surgery where blood supply to the liver is temporary interrupted. IRI causes up to 10% of early transplant failures and can lead to significantly higher incidence of acute and chronic rejections. Thousands of patients die every year while waiting for a donor organ. This gloomy scenario, together with the significant prevalence of obesity in the population, has led to an increased need of using suboptimal steatotic livers in transplantation at elevated risks of dysfunction based on their high susceptibility to IRI. In the previous funding period, our results demonstrated that specific matrix metalloproteinase-9 (MMP-9) inhibition profoundly ameliorates IRI in normal livers, and unveiled potential key roles for MMP-2 and TIMP-1 in liver injury. This proposal (i) explores the hepatoprotective properties of MMP-9 inhibition in marginal steatotic liver IRI and (ii) dissects the functions of MMP-2 and TIMP-1 in IR-induced damage in both normal and steatotic livers. We expect that the proposed work will provide fundamental insights on the individual functions of key MMPs/TIMPs, leading to the development of novel therapeutic manipulations that can minimize their detrimental effects while maximizing their beneficial functions in normal and in steatotic liver IRI. Well-established models of partial liver IRI in normal and in steatotic mice, and of ex vivo cold steatotic liver ischemia followed by iso-transplantation in rats will be used to address the following aims: (1) To dissect mechanisms by which Matrix Metalloproteinase-9 specific inhibition affects the IRI outcome in marginal steatotic livers. Using MMP-9-/- deficient mice and specific therapeutic manipulations against MMP-9, we will dissect whether specific MMP-9 inhibition (i) protects marginal steatotic livers from the I/R insult, (ii) disrupts leukocyte traffic and chemokine release/activation, and we will (iii) identify intracellular signaling mechanisms leading to MMP-9 expression in steatotic hepatic IRI; (2) To analyze mechanisms by which Matrix Metalloproteinase-2 activity affects the IRI outcome in normal and in steatotic livers. We will determine whether selective MMP-2 inhibition (i) affects liver function/preservation, (ii) results in altered expression of MMP-9, (iii) interferes with patterns of leukocyte migration and of cytokine/chemokine activation and, furthermore, we will (iv) identify the sources of MMP- 2 and their relative contribution in normal and steatotic liver IRI; and (3) To dissect the mechanisms by which tissue inhibitor of metalloproteinases-1 affects IRI in normal and in steatotic livers. We will assess the function of TIMP-1 in normal and in steatotic liver IRI by determining the impact of TIMP-1 deficiency on (i) liver function/preservation, (ii) liver regeneration and apoptosis, (iii) MMP activation, and on (iii) leukocyte recruitment and pro-inflammatory networks. Moreover, we will assess the function of Timp-1 overexpression as a therapeutic approach in partial liver IRI and in steatotic OLT.

肝缺血再灌注损伤（IRI）发生在所有移植肝、创伤、休克和择期手术中。 肝脏血液供应暂时中断的手术。IRI导致高达10%的早期移植 失败，并可能导致急性和慢性排斥反应的发生率显着较高。成千上万的 每年都有病人在等待捐赠器官时死亡。这一令人沮丧的情况，加上 肥胖症在人群中的显著流行，已经导致使用次优的 脂肪变性肝在移植中的功能障碍风险增加，因为它们对IRI的高度易感性。 在上一个资助期，我们的研究结果表明，特异性基质金属蛋白酶-9（MMP-9） 抑制显著改善正常肝脏的IRI，并揭示了MMP-2的潜在关键作用， TIMP-1在肝损伤中的作用该建议（i）探索MMP-9抑制在肝细胞中的肝保护特性。 边缘性脂肪肝IRI和（ii）解剖MMP-2和TIMP-1在IR诱导的损伤中的功能， 正常和脂肪变性的肝脏我们希望拟议的工作将提供基本的见解， 关键MMPs/TIMPs的个体功能，导致新的治疗药物的开发， 操作，可以最大限度地减少其有害影响，同时最大限度地发挥其有益功能， 正常和脂肪肝IRI。在正常和脂肪变性中建立了部分肝脏IRI的良好模型 小鼠，以及离体冷脂肪变性肝缺血后在大鼠中进行同种移植， 解决以下目标：（1）剖析基质金属蛋白酶-9特异性 抑制影响边缘脂肪变性肝脏的IRI结果。使用MMP-9-/-缺陷小鼠和特异性 针对MMP-9的治疗操作，我们将分析特异性MMP-9抑制是否（i）保护 来自I/R损伤的边缘性脂肪肝，（ii）破坏白细胞运输和趋化因子释放/活化，和 我们将（iii）确定导致脂肪变性肝IRI中MMP-9表达的细胞内信号传导机制;（2） 分析基质金属蛋白酶-2活性影响正常人IRI结局的机制， 和脂肪肝中。我们将确定选择性MMP-2抑制（i）是否影响肝脏 功能/保存，（ii）导致MMP-9的表达改变，（iii）干扰白细胞的模式 迁移和细胞因子/趋化因子激活，此外，我们将（iv）确定MMP-1的来源， 2及其在正常和脂肪肝IRI中的相对贡献;（3）通过以下方法剖析其机制： 金属蛋白酶-1的组织抑制剂影响正常肝脏和脂肪变性肝脏的IRI。我们将 通过测定TIMP-1的影响，评估TIMP-1在正常和脂肪肝IRI中的功能 （i）肝功能/保存，（ii）肝再生和凋亡，（iii）MMP活化，和 （iii）白细胞募集和促炎网络。此外，我们将评估 Timp-1过表达作为部分肝脏IRI和脂肪变性奥尔特的治疗方法