Protective Effect of Hemin Preconditioning after Intracerebral Hemorrhage

氯化血红素预处理对脑出血后的保护作用

基本信息

批准号：
8346310
负责人：
RAYMOND F REGAN
金额：
$ 33.91万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8346310
关键词：
Accounting Acute Intermittent Porphyria Adhesives Aftercare Anti-Inflammatory Agents Anti-inflammatory Antibodies Antioxidants Astrocytes Attenuated Autologous Behavioral Blood Blood - brain barrier anatomy Blood Clot Blood Vessels Blood coagulation Brain Brain Injuries Brain hemorrhage Cell Adhesion Molecules Cell Death Cell Survival Cells Cerebral hemisphere hemorrhage Clinical Clinical Trials Coagulation Process Cognitive deficits Complex Corpus striatum structure Cytolysis Deposition Development Disease Dose Edema Endothelial Cells Enzymes Erythrocytes Event Excision Functional disorder Gene Transfer Goals Harvest Hematoma Heme Hemin Hemoglobin Hemopexin Hemorrhage Home environment Hour Inflammation Inflammatory Inflammatory Infiltrate Injection of therapeutic agent Injury Intraperitoneal Injections Iron Ischemia Ischemic Stroke Knock-out Knockout Mice Laboratories Lead Lesion Life Mediating Microglia Modeling Morbidity - disease rate Mus Nature Nerve Degeneration Neuroglia Neurologic Neurons Oligodendroglia Outcome Oxygenases Peripheral Population Prevalence Process Protein Isoforms Proteins Publishing Resistance Safety Serine Protease Stimulus Stroke Survivors Testing Therapeutic Thrombin Time Tissues Toxic effect Toxin Transgenic Mice Traumatic Brain Injury Video Recording Wild Type Mouse cell injury collagenase digital effective therapy heme oxygenase-1 heme-binding protein improved interest intraperitoneal mortality novel object recognition overexpression oxidation preconditioning protective effect protein expression research study response

项目摘要

DESCRIPTION (provided by applicant): Intracerebral hemorrhage (ICH) is the primary event in 10-15% of strokes. A growing body of experimental evidence supports the hypothesis that release of hemin from the hematoma may contribute to oxidative cell injury in adjacent tissue. The heme oxygenase (HO) enzymes, which catalyze the rate-limiting step in hemin breakdown, regulate the response of CNS cells to hemin in a complex fashion. HO-1, the inducible isoform, has been associated with increased lesion volume and inflammation after experimental ICH, and worsened behavioral outcome. However, recent studies in the applicant's laboratory suggest that mice overexpressing HO-1 specifically in astrocytes are markedly less vulnerable to ICH than wild-type mice. Moreover, systemic administration of hemin, which is currently in clinical use to treat acute porphyrias, increased HO-1 expression in the mouse brain. When administered after ICH but before erythrocyte lysis, this treatment attenuated blood- brain barrier breakdown and cell injury surrounding a striatal hematoma. These results suggest that systemic hemin at clinically-tolerated doses provides a preconditioning stimulus that protects CNS cells from the hemin subsequently released in high concentrations from the hematoma. This safe, easily administered, and inexpensive compound may be a novel and highly effective treatment for ICH. The goal of this project is to define the therapeutic benefit of systemic hemin in two established ICH models. The specific aims are as follows: 1) Administer hemin to mice via daily i.p. injections for 1-3 days; 24 hours after the last injection, harvest striata and quanify HO-1 protein expression and activity. Identify cell populations that express HO-1 via immunostaining. 2) Induce ICH by stereotactic injection of autologous blood or collagenase into the striata of mice. Treat with hemin or vehicle control 1, 3, 6, or 12 hours later, followed in 24 hours by an additional dose. Quantify blood-brain barrier disruption and striatal edema at 3 days. 3) Quantify the effect of hemin on striatal cell viability and perihematomal inflammatory infiltrates 3 and 8 days after ICH. Assess focal deficits at these time points using corner, adhesive removal and elevated body swing tests, and activity deficits by digital analysis of home cage video recordings. 4) Compare the effects of hemin treatment on blood-brain barrier disruption, edema, neuronal viability, inflammation, and behavioral deficits in wild-type mice with those in HO-1 knockout mice. Determine the effect of the heme binding protein hemopexin on hemin therapy by performing additional experiments using hemopexin knockout mice. It is hoped that the results of this project will define the benefits of systemic hemin after ICH, and rapidly lead to clinical trials for a disease process that currently has few therapeutic options. PUBLIC HEALTH RELEVANCE: The information gained in this project may lead to new treatments for victims of hemorrhagic brain injuries. These include hemorrhagic stroke and traumatic brain injuries associated with bleeding into the brain. The ultimate goal is to reduce the cell injury that occurs in tissue surrounding the blood clot, and thereby improve the likelihoo of survival and return to an independent, productive life.

描述（由申请人提供）：脑出血 (ICH) 是 10-15% 中风的主要事件。越来越多的实验证据支持这样的假设：血肿中释放的血红素可能导致邻近组织的氧化细胞损伤。血红素加氧酶 (HO) 催化血红素分解的限速步骤，以复杂的方式调节中枢神经系统细胞对血红素的反应。 HO-1 是一种诱导型亚型，与实验性 ICH 后病变体积和炎症的增加以及行为结果恶化有关。然而，申请人实验室的最新研究表明，与野生型小鼠相比，在星形胶质细胞中特异性过度表达HO-1的小鼠明显不易受到ICH的影响。此外，目前临床上用于治疗急性卟啉症的氯高铁血红素的全身给药可增加小鼠大脑中 HO-1 的表达。当 ICH 之后、红细胞裂解之前进行治疗时，这种治疗可以减轻血脑屏障的破坏和纹状体血肿周围的细胞损伤。这些结果表明，临床耐受剂量的全身血红素提供了一种预处理刺激，可以保护中枢神经系统细胞免受随后从血肿释放的高浓度血红素的影响。这种安全、易于给药且廉价的化合物可能是一种新颖且高效的脑出血治疗方法。该项目的目标是在两个已建立的 ICH 模型中确定全身血红素的治疗益处。具体目标如下： 1）每天通过腹膜内注射给予小鼠血红素。注射1-3天；最后一次注射后 24 小时，收获纹状体并量化 HO-1 蛋白表达和活性。通过免疫染色鉴定表达 HO-1 的细胞群。 2）通过向小鼠纹状体立体定向注射自体血液或胶原酶来诱导ICH。 1、3、6 或 12 小时后用氯高铁血红素或溶媒对照进行治疗，24 小时后进行治疗小时额外剂量。量化第 3 天的血脑屏障破坏和纹状体水肿。 3) 量化 ICH 后 3 天和 8 天血红素对纹状体细胞活力和血肿周围炎症浸润的影响。使用角落、粘合剂去除和升高的身体摆动测试来评估这些时间点的焦点缺陷，并通过对家庭笼子视频记录的数字分析来评估活动缺陷。 4) 比较血红素治疗对野生型小鼠的血脑屏障破坏、水肿、神经元活力、炎症和行为缺陷的影响 HO-1 敲除小鼠中的那些。通过使用血红素结合蛋白敲除小鼠进行额外的实验，确定血红素结合蛋白血红素治疗对血红素治疗的影响。希望该项目的结果将明确 ICH 后全身血红素的益处，并迅速开展针对目前几乎没有治疗选择的疾病过程的临床试验。公共卫生相关性：该项目中获得的信息可能会为出血性脑损伤患者带来新的治疗方法。这些包括出血性中风和与脑出血相关的创伤性脑损伤。最终目标是减少血凝块周围组织中发生的细胞损伤，从而提高生存并恢复独立、富有成效的生活的可能性。