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）是由于血流中断而引起的新生儿大脑损伤。 1000 名足月新生儿中的 2-4 名和 60% 的早产儿是死亡率和发病率的主要原因。 与终生神经功能障碍有关。 内质网 (ER) 应激是 HI 后遇到的主要病理，与内质网失调相关 导致细胞凋亡和炎症的蛋白质折叠 HI 诱导的 ER 应激上调促细胞凋亡。 肌醇需要酶 1 α (IRE1α) 信号通路，也与活性氧相关 (ROS) 积累，主要来自 NADPH 依赖性细胞色素 P450 还原酶 (NPR) 和 P450 2E1 （CYP）复合物。 Bax 抑制剂 1 (BI-1) 蛋白表达于 ER 膜上，已被证明在抑制 ER 中发挥重要作用 应激诱导的信号通路可以直接与 IRE1α 结合，从而抑制这一促凋亡通路。 以及通过解离 NPR-CYP 复合物来减少 ROS 积累。 本研究的目的是确定 BI-1 在体外氧气中的抗凋亡和抗炎作用 葡萄糖剥夺 (OGD) 模型和体内新生 HI 大鼠模型以及阐明机制 我们的中心假设是（1）用Ad-转染细胞。 TMBIM6 载体将提高 OGD 后的细胞活力，并帮助确定 BI-s-1 主要途径 (2)； 通过 Ad-TMBIM6 注射在大脑中过度表达 BI-1 蛋白将改善新生儿 HI 后的恢复 通过减少 ER 诱导的应激 (a) 通过抑制 IRE1α 信号通路来减少神经元凋亡，以及 (b) 通过 NPR-CYP 复合物的解离和随后的 ROS 抑制而引起神经炎症。 具体目标 1： 确定 ER 应激信号通路在抗凋亡和抗炎机制中的作用 BI-1 在体外缺氧葡萄糖剥夺 (OGD) 模型中的作用 具体目标 2：确定 BI-1 是否存在。 在体内新生 HI 大鼠模型中，上调通过 IRE1α 信号通路发挥抗凋亡作用。 具体目标3：研究BI-1过表​​达的抗炎作用及其信号通路 参与体内新生 HI 大鼠模型。 该提案的长期目标是： 1) 建立 BI-1 作为 ER 应激的主要调节器 2) 建立 BI-1 新生儿HI后的信号通路；3)为BI-1作为潜在治疗靶点提供基础。