Mitochondrial Carbonic Anhydrases and Diabetic Blood-Brain Barrier Disruption

线粒体碳酸酐酶和糖尿病血脑屏障破坏

基本信息

批准号：
8604388
负责人：
WILLIAM A BANKS
金额：
$ 32.1万
依托单位：
SAINT LOUIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-02-01 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8604388
关键词：
Acetyl Coenzyme A Affect Apoptosis Astrocytes Bicarbonates Binding Blood Blood - brain barrier anatomy Blood Vessels Blood-Retinal Barrier Brain Carbonic Anhydrase Inhibitors Catabolism Cell Respiration Cells Citric Acid Citric Acid Cycle Coculture Techniques Communication Complications of Diabetes Mellitus Cytoplasm Deterioration Development Diabetes Mellitus Drug usage Electron Transport Endothelial Cells Extravasation Functional disorder Generations Glucose Hexosamines Hyperglycemia In Vitro Insulin Insulin Resistance Knock-out Laboratories Lead Marketing Measures Membrane Metabolism Methods Mitochondria Modeling Mus Nervous System Physiology Neuraxis Oxaloacetates Oxidative Stress Pathway interactions Pericytes Pharmaceutical Preparations Plasma Proteins Process Production Property Pyruvate Reactive Oxygen Species Resistance Retinal Role Serum Proteins Source Streptozocin System Testing Toxic effect Translational Research Wild Type Mouse Work aerobic glycolysis carbonate dehydratase cell type diabetic glucose metabolism in vitro Model in vivo in vivo Model inhibitor/antagonist mitochondrial membrane neurovascular unit overexpression polyol prevent public health relevance research study

项目摘要

DESCRIPTION (provided by applicant): Diabetes mellitus is associated with deterioration of the brain's microvasculature and the blood-brain barrier (BBB) that it forms. The BBB prevents the unrestricted leakage of plasma proteins into the brain. The vascular BBB is physically formed by specially modified brain endothelial cells (BECs), but other cell types interact with it to form the neurovascular unit (NVU). Pericytes in particular are important in maintaining BBB function in the face of glycemic insult. We have shown in vitro that the insulin transporter is lost in the face of high glucose concentrations unless BECs are co-cultured with pericytes; astrocyte co-culture does not preserve BEC insulin transporter function. However, pericytes are themselves susceptible to glucose toxicity and the accompanying oxidative stress. Work with retinal pericytes and the blood-retinal barrier (BRB) shows that hyperglycemia induces apoptosis in retinal pericytes; with pericyte loss the BRB deteriorates. Mitochondrial carbonic anhydrases (CAs) are important in the generation of reactive oxygen species (ROS) and the subsequent oxidative stress that arises and is accelerated during hyperglycemia. In brief, mitochondrial CAs generate mitochondrial bicarbonate that is necessary for the oxidative catabolism of glucose, the major source of ROS. Hyperglycemia accelerates this process, increasing production of ROS. Blocking mitochondrial CAs shuttles pyruvate through anaerobic metabolism and so prevents excessive ROS production. We hypothesize that inhibition of mitochondrial carbonic anhydrases (CAs) will protect pericytes and BECs from ROS induced apoptosis and slow the development of disruption of the BBB in STZ-induced diabetes. Wewill test this hypothesis in two specific aims. SA1 will measure oxidative stress induced by high glucose media and the resulting mitochondrial leakage and apoptosis in primary cultures of brain pericytes and BEC isolated from mitochondrial CA KO & WT mice and in pericytes and BEC that overexpress CA or are treated with clinically used CA inhibitors. SA 2 will measure in diabetic and control mice BBB disruption and oxidative stress in BEC from CA KO mice, WT mice, and mice treated with the CA inhibitors of SA1.

描述（由申请人提供）：糖尿病与大脑微脉管系统的恶化和形成的血脑屏障（BBB）有关。 BBB防止血浆蛋白不受限制地渗入大脑。血管BBB是由特殊修饰的脑内皮细胞（BEC）物理形成的，但其他细胞类型与之相互作用以形成神经血管单元（NVU）。尤其是周细胞在面对血糖损伤时保持BBB功能很重要。我们已经在体外表明，除非BEC与周细胞共培养，否则胰岛素转运蛋白会损失。星形胶质细胞共培养不保留BEC胰岛素转运蛋白的功能。但是，周细胞本身容易受到葡萄糖毒性和随附的氧化应激。与视网膜周期和血液视网膜屏障（BRB）一起工作表明，高血糖会诱导视网膜周细胞细胞凋亡。随着周细胞的损失，BRB恶化。线粒体碳酸酐酶（CAS）在活性氧（ROS）的产生中很重要，并且在高血糖期间产生并加速了随后的氧化应激。简而言之，线粒体CAS产生线粒体碳酸氢盐，这对于葡萄糖的氧化分解代谢是ROS的主要来源。高血糖加速了这一过程，增加了ROS的产生。通过厌氧代谢阻止线粒体CAS穿梭丙酮酸，因此可以防止过度的ROS产生。我们假设抑制线粒体碳酸酐酶（CAS）将保护周细胞和BEC免受ROS诱导的细胞凋亡，并减缓BBB在STZ诱导的糖尿病中的破坏。我们将以两个具体的目的检验这一假设。 SA1将测量由高葡萄糖培养基诱导的氧化应激，并在脑周细胞的原发性培养物中产生的线粒体泄漏和凋亡，并从线粒体CA KO＆WT小鼠和周细胞中分离出来，并且在周细胞和周围和临床中过表达CA或用临床使用的CA抑制剂处理过表达。 SA 2将在CA KO小鼠，WT小鼠和用SA1的CA抑制剂治疗的CA KO小鼠，WT小鼠和小鼠的BEC中测量糖尿病和对照小鼠的BBB破坏和氧化应激。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Topiramate Protects Pericytes from Glucotoxicity: Role for Mitochondrial CA VA in Cerebromicrovascular Disease in Diabetes.

托吡酯保护周细胞免受糖毒性：线粒体 CA VA 在糖尿病脑微血管疾病中的作用。

DOI：
10.15226/2374-6890/2/2/00123
发表时间：
2015
期刊：
Journal of endocrinology and diabetes
影响因子：
0
作者：
Patrick,Ping;Price,TulinO;Diogo,AnaL;Sheibani,Nader;Banks,WilliamA;Shah,GulN
通讯作者：
Shah,GulN

High glucose-induced mitochondrial respiration and reactive oxygen species in mouse cerebral pericytes is reversed by pharmacological inhibition of mitochondrial carbonic anhydrases: Implications for cerebral microvascular disease in diabetes.

DOI：
10.1016/j.bbrc.2013.09.086
发表时间：
2013-10-18
期刊：
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
影响因子：
3.1
作者：
Shah, Gul N.;Morofuji, Yoichi;Banks, William A.;Price, Tulin O.
通讯作者：
Price, Tulin O.

Protective Effect of Topiramate on Hyperglycemia-Induced Cerebral Oxidative Stress, Pericyte Loss and Learning Behavior in Diabetic Mice.

托吡酯对糖尿病小鼠高血糖引起的脑氧化应激、周细胞损失和学习行为的保护作用。

DOI：
发表时间：
2015
期刊：
International library of diabetes & metabolism
影响因子：
0
作者：
Price,TulinO;Farr,SusanA;Niehoff,MichaelL;Ercal,Nuran;Morley,JohnE;Shah,GulN
通讯作者：
Shah,GulN