Endothelial-Pericyte Crosstalk in Diabetic Stroke

糖尿病中风的内皮-周细胞串扰

基本信息

批准号：
10338161
负责人：
Nabil J Alkayed
金额：
$ 41.75万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10338161
关键词：
Acute Alteplase Blood - brain barrier anatomy Blood Platelets Blood Vessels Blood capillaries Blood flow Brain Caliber Capillary Endothelial Cell Cells Cerebrovascular Circulation Cerebrum Clinical Coagulation Process Cytolysis Diabetes Mellitus Diabetic mouse Edema Eicosanoids Electron Microscopy Endothelium Enzymes Epoxide hydrolase Excision FDA approved Failure Fibrin Filament G-Protein-Coupled Receptors GPR39 gene GTP-Binding Protein alpha Subunits, Gs Glucose Hemorrhage High Fat Diet Histology Immunohistochemistry In Situ In Vitro Infarction Injections Injury Ischemia Ischemic Stroke Knockout Mice Leukocytes Link Lipids Measurement Mechanics Mediating Methods Middle Cerebral Artery Occlusion Morphology Mus Neurocognitive Niacinamide Non-Insulin-Dependent Diabetes Mellitus Obstruction Optics Outcome Oxygen Pathogenesis Patients Perfusion Pericytes Pharmaceutical Preparations Pharmacology Platelet Activation Platelet aggregation Prevention Reperfusion Therapy Role Signal Transduction Streptozocin Stroke Swelling Testing Thrombectomy Thrombin Thrombus Time Tissues Transgenic Mice Transgenic Organisms Type 2 diabetic Vasodilator Agents alpha-Thrombin blood-brain barrier disruption brain tissue cell type cerebrovascular constriction deprivation diabetic functional independence improved in vivo intravital imaging ischemic injury leukocyte activation microangiography middle cerebral artery neurobehavioral novel overexpression post stroke preservation prevent protective effect receptor restoration somatosensory stroke model stroke outcome stroke therapy thromboembolic stroke two photon microscopy

项目摘要

Project Summary Tissue plasminogen activator (tPA) remains the only FDA-approved therapy for acute ischemic stroke. Unfortunately, one-third to half of patients with successful recanalization of large cerebral vessels do not have good clinical outcome. This is in part due to incomplete microvascular reperfusion termed focal “no-reflow”, which is particularly prevalent in diabetic stroke. The current proposal will determine the role of endothelial-pericyte crosstalk, mediated by the action of the endothelial eicosanoids epoxyeicosatrienoates (EETs) on pericytes, in microvascular no-reflow after diabetic stroke in mice. We will test the hypothesis that endothelial-derived EETs preserve capillary perfusion in brain after ischemia, in part by protecting pericytes from ischemic injury, and that diabetes reduces microvascular endothelial EETs; thus, leading to greater pericyte injury and capillary damage after diabetic stroke. We will also determine if EETs protect pericytes by inhibiting G-protein coupled receptor 39 (GPR39). Stroke is induced in mice with and without type 2 diabetic (T2D), and with higher or lower endothelial EETs (due to transgenic overexpression or deletion of EETs-degrading enzyme soluble epoxide hydrolase, sEH, in endothelium) using an intraluminal occlusive filament or in-situ thrombin injection to occlude the middle cerebral artery (MCA). Intra-vital imaging using optical microangiography (OMAG) and two-photon microscopy (2PM) will be used to assess pericyte morphology, BBB integrity, capillary blood flow, and platelet and leukocyte morphology and dynamics within the peri-infarct region in-vivo, in real-time after MCA occlusion. Mice will be survived for 4 days after stroke to assess cellular and tissue damage (H&E, EM, IHC), and for 28 days to assess long-term functional deficit (neurocognitive and somatosensory). We propose that preserving endothelial- pericyte EETs/GPR39 signaling protects pericytes, prevents no-reflow and reduces brain tissue damage and neurobehavioral functional deficit. Enhancing endothelial EETs, either pharmacologically or in transgenic mice with higher endothelial EETs, will also enhance tPA efficacy and reduce tPA-associated hemorrhage after stroke in diabetic mice.

项目概要组织纤溶酶原激活剂（tPA）仍然是 FDA 批准的唯一治疗急性缺血性中风的疗法。不幸的是，三分之一到一半的大脑血管成功再通的患者没有良好的临床结果部分归因于称为局部“无复流”的不完全微血管再灌注。在糖尿病中风中尤其常见，当前的提议将确定内皮周细胞的作用。串扰，由内皮类二十烷酸环氧二十碳三烯酸酯 (EET) 对周细胞的作用介导，小鼠糖尿病中风后微血管无复流我们将检验内皮源性 EET 的假设。保护缺血后大脑中的毛细血管灌注，部分是通过保护周细胞免受缺血性损伤，并且糖尿病会减少微血管内皮 EET，从而导致更大的周细胞损伤和毛细血管损伤；我们还将确定 EET 是否通过抑制 G 蛋白偶联受体 39 来保护周细胞。 (GPR39) 在患有或不患有 2 型糖尿病 (T2D) 以及具有较高或较低内皮细胞的小鼠中诱发中风。 EET（由于 EET 降解酶可溶性环氧化物水解酶、sEH 的转基因过度表达或缺失，在内皮细胞中）使用腔内闭塞丝或原位凝血酶注射来闭塞中间层使用光学微血管造影 (OMAG) 和双光子显微镜进行脑动脉 (MCA) 活体成像。 (2PM) 将用于评估周细胞形态、血脑屏障完整性、毛细血管血流量以及血小板和白细胞小鼠大脑中动脉 (MCA) 闭塞后，实时观察梗塞周围区域的形态和动态。中风后存活 4 天以评估细胞和组织损伤（H&E、EM、IHC），并存活 28 天以评估我们建议保留内皮细胞的长期功能缺陷（神经认知和体感）。周细胞 EET/GPR39 信号传导可保护周细胞，防止无复流并减少脑组织损伤通过药理学或转基因小鼠增强内皮 EET。具有更高的内皮 EET，还将增强 tPA 功效并减少中风后与 tPA 相关的出血在糖尿病小鼠中。