ER stress and neonatal hypoxia ischemia encephalopathy

内质网应激与新生儿缺氧缺血性脑病

• 批准号: 10059275
• 负责人: John H Zhang
• 金额: $ 34.56万
• 依托单位: LOMA LINDA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10059275
• 关键词: Affect; Animals; Anti-Inflammatory Agents; Antiinflammatory Effect; Apoptosis; Apoptotic; Attenuated; BCL2 gene; Binding; Blood; Blood flow; Brain; Brain Hypoxia-Ischemia; Brain Injuries; Cause of Death; Cell Survival; Cells; Clinical; Complex; Cytochrome P450; Disease; Dissociation; Electron Transport; Emotional; Encephalopathies; Endoplasmic Reticulum; Enzymes; Generations; Genes; Glucose; Goals; Human; Hypoxic-Ischemic Brain Injury; Impairment; In Vitro; Infant; Inflammation; Injections; Injury; Inositol; Interruption; Life; Link; Long-Term Effects; Medical; Membrane; Modeling; Morbidity - disease rate; Morphology; NADP; NADPH-Ferrihemoprotein Reductase; Neurologic; Neurological outcome; Oxygen; PC12 Cells; Pathology; Pathway interactions; Play; Premature Infant; Process; Property; Protein Family; Proteins; Rattus; Reactive Inhibition; Reactive Oxygen Species; Recovery; Role; Signal Pathway; Signal Transduction; Stress; Survivors; Term Birth; Testing; Therapeutic; Transfection; Up-Regulation; attenuation; deprivation; endoplasmic reticulum stress; improved; in vivo; inhibitor/antagonist; mortality; neonatal brain; neonatal hypoxic-ischemic brain injury; neuroinflammation; neuron apoptosis; neuroprotection; overexpression; protective effect; protein folding; sensor; social; stroke model; therapeutic target; transcription factor; vector

Neonatal hypoxia ischemia (HI) is an injury to the neonatal brain caused by interrupted blood flow. It occurs in 2-4 of 1000 full-term births and 60% of premature infants. It is the leading cause of mortality and morbidity associated with life-long neurological impairments. Endoplasmic reticulum (ER) stress is a major pathology encountered after HI, associated with dysregulation of protein folding leading to apoptosis and inflammation. HI induced ER stress up regulates the pro-apoptotic Inositol requiring enzyme-1 alpha (IRE1α) signaling pathway and is also associated with reactive oxygen species (ROS) accumulation, mainly from the NADPH-dependent cytochrome P450 reductase (NPR) and P450 2E1 (CYP) complex. Bax-inhibitor 1 (BI-1) protein, expressed on ER membrane, has been shown to play a major role in inhibiting ER stress induced signaling pathways. BI-1 can directly bind to IRE1α thus inhibiting this pro-apoptotic pathway as well as reduce ROS accumulation by dissociating the NPR-CYP complex. The objective of this study is to establish BI-1s anti-apoptotic and anti-inflammatory effects in an in vitro oxygen glucose deprivation (OGD) model and in an in vivo neonatal HI rat model as well as to elucidate the mechanisms via which it confers its protective properties. Our central hypothesis is that (1) transfection of cells with Ad- TMBIM6 vector will improve cell viability after OGD as well as help determine BI-s-1s major pathways; (2) overexpression of the BI-1 protein in the brain, via Ad-TMBIM6 injection will improve recovery after neonatal HI by reducing ER stress induced (a) neuronal apoptosis via inhibition of IRE1α signaling pathway and (b) neuroinflammation via dissociation of the NPR-CYP complex and subsequent inhibition of ROS. Specific Aim 1: To determine the role of ER stress signaling pathways in the anti-apoptotic and anti-inflammatory mechanisms of BI-1 in an in vitro Oxygen Glucose Deprivation (OGD) model. Specific Aim 2: To determine whether BI-1 upregulation exerts its anti-apoptotic effects via the IRE1α signaling pathway in an in vivo neonatal HI rat model. Specific Aim 3: To investigate the anti-inflammatory effects of BI-1 overexpression and the signalling pathways involved in an in vivo neonatal HI rat model. The long-term goals of this proposal are to: 1) establish BI-1 as main regulator of ER stress 2) establish BI-1s signaling pathways after neonatal HI; 3) provide a basis for BI-1 as a potential therapeutic target.

新生儿缺氧缺血(HI)是由于血流中断对新生儿脑造成的损伤。它发生在 每1000名足月新生儿中有2-4名，早产儿中有60%。它是导致死亡和发病的主要原因。 与终生神经损伤有关。 内质网应激是缺氧缺血性脑损伤后的主要病理改变，与血管内皮细胞功能紊乱有关。 蛋白质折叠导致细胞凋亡和炎症。Hi诱导的内质网应激上调促细胞凋亡 肌醇需要酶-1α(IRE1-α)信号通路，并与活性氧有关 (ROS)积累，主要来自NADPH依赖的细胞色素P450还原酶(NPR)和P450 2E1 (CYP)情结。 Bax-Inhibitor 1(BI-1)蛋白表达于内质网，在抑制内质网中起重要作用 应激诱导的信号通路。BI-1可以直接与IRE1α结合，从而抑制这一促凋亡途径，如 并通过解离NPR-CYP复合体来减少ROS的积累。 本研究的目的是在体外氧气中建立BI-1的抗细胞凋亡和抗炎作用。 葡萄糖剥夺(OGD)模型和在体新生HI大鼠模型及其机制的研究 通过它赋予它的保护属性。我们的中心假设是(1)将Ad-1基因导入细胞。 TMBIM6载体将提高OGD后的细胞存活率，并有助于确定BI-S-1s的主要途径；(2) 通过注射Ad-TMBIM6在脑内过表达BI-1蛋白将促进新生儿缺氧缺血性脑损伤后的恢复 减少内质网应激诱导的神经元凋亡(A)通过抑制IRE1α信号通路和(B) 通过NPR-CYP复合体的解离和随后的ROS抑制而引起的神经炎症。具体目标1： 确定内质网应激信号通路在抗细胞凋亡和抗炎机制中的作用 BI-1在体外氧糖剥夺(OGD)模型中的表达。具体目标2：确定BI-1是否 上调通过IRE1α信号通路在体内新生HI大鼠模型中发挥抗凋亡作用。 特异性目标3：探讨BI-1过表达的抗炎作用及其信号转导途径 建立新生大鼠缺氧缺血性脑病动物模型。 这项建议的长期目标是：1)建立BI-1作为内质网应激的主要调节因子2)建立BI-1 3)为BI-1作为潜在的治疗靶点提供了依据。