Peptide therapy for alcohol-induced CNS injury

摘要 酒精是美国使用和滥用最多的毒品。慢性酒精滥用导致神经元 感觉运动、记忆、心理和认知功能的退化和功能缺陷。我们和 其他人已经证明，氧化信号是酒精诱导的损伤的中心机制，导致 神经和功能缺陷。修复积累的氧化自由基可作为一种 预防酗酒者神经损害进展的有效策略。因此，这项提议将 研究脑损伤及其相关的神经炎症和神经退行性变的新假说 由酒精引起的氧化损伤可以通过激活抗氧化信号Nrf2来修复 (核因子E2相关因子2)途径。Nrf2转录系统是一种内源性防御系统 通过以下途径促进许多解毒、细胞保护和抗炎基因的表达 与其调控区域中的抗氧化反应元件(ARE)相互作用。NRF2有可能成为 一个新的治疗靶点，以减轻酒精引起的神经损伤。我们的中心假设是 抗氧化剂促进转录因子Nrf2的激活减轻氧化应激诱导 与过量饮酒有关的病理生理变化。这些更改包括激活 转化生长因子β-1和基质金属蛋白酶-1与神经炎性反应， 和神经退化。Nrf2转录因子的激活与内源性抗氧化剂的上调 用Nrf2激活剂III TAT多肽(Nrf2多肽)治疗动物将获得酶。我们会 系统地(皮下)给酒精摄入小鼠注射Nrf2多肽及其疗效评价 前额叶等特定区域减轻酒精相关神经损害的潜力 大脑皮层、海马体和小脑。拟议的研究不仅验证了NRF2的因果关系 多肽在缓解AUD氧化应激中的作用，但也识别了Nrf2转录的机制调节 用芯片定量聚合酶链式反应和芯片序列分析研究了因子激活的抗氧化剂基因及其与Nrf2的相互作用。这个 Nrf2的保护作用将通过评估其对基因表达和蛋白质水平的影响来确定。 主要的抗氧化剂、炎症和凋亡蛋白。我们将验证Nrf2肽在Nrf2中的作用 基因敲除(KO)小鼠(NRF2−/−)。我们还将使用一组行为测试来评估认知、 与Nrf2肽治疗相关的感觉-运动反馈和心理行为恢复 在野生型和Nrf2-/-小鼠中进行酒精暴露。我们将结果与对照多肽进行比较。 (带有TAT的随机序列)处理的动物。我们预计这项研究的结果将会有 广泛的临床相关性。

ABSTRACT Alcohol is the most used and abused drug in the United States. Chronic alcohol abuse results in neuronal degeneration and functional deficits in sensorimotor, memory, psychological, and cognitive functions. We and others have demonstrated that oxidative signaling is the central mechanism in alcohol-induced injury that leads to neurological and functional deficits. Remediation of accumulating oxidative radicals may serve as an effective strategy for preventing the progression of neurological damage in alcoholics. Thus, this proposal will study a novel hypothesis that the brain injury and associated neuroinflammation and neurodegeneration caused by alcohol-induced oxidative damage can be repaired by activating the antioxidant signaling Nrf2 (nuclear factor E2-related factor 2) pathway. The Nrf2 transcriptional system is an endogenous defense mechanism that boosts the expression of many detoxifying, cytoprotective and anti-inflammatory genes by interacting with the antioxidant response element (ARE) in their regulatory regions. Nrf2 has the potential to be a novel and therapeutic target to mitigate alcohol-induced neural damage. Our central hypothesis is that activation of antioxidant-promoting transcription factor Nrf2 alleviates oxidative stress-induced pathophysiological changes associated with excessive alcohol ingestion. These changes include activation of transforming growth factor β1 (TGF-β1) and matrix metalloproteinases (MMPs), neuroinflammatory responses, and neurodegeneration. Activation of Nrf2 transcription factor and up-regulation of endogenous antioxidant enzymes will be achieved by treating animals with Nrf2 activator III TAT peptide (Nrf2 peptide). We will administer Nrf2 peptide to the alcohol ingested mice systemically (subcutaneous) and evaluate its therapeutic potential in alleviating alcohol-associated neurological impairments in defined regions such as the prefrontal cortex, hippocampus, and cerebellum. The proposed study not only validates the cause and effect of Nrf2 peptide in mitigating oxidative stress in AUD but also identifies the mechanistic regulation of Nrf2 transcription factor activated antioxidant genes and their interactions with Nrf2 by ChIP-qPCR, and ChIP-Seq. The protective role of Nrf2 will be determined by assessing its effects on gene expression and protein levels of the major antioxidant, inflammatory, and apoptotic proteins. We will validate the effect of Nrf2 peptide in Nrf2 knock-out (KO) mice (Nrf2−/−). We will also use a cohort of behavioral tests that will utilize to assess cognitive, sensory-motor feedback, and psychological behavioral recovery associated with Nrf2 peptide treatment following alcohol exposure in wild type and Nrf2-/- mice. We will compare the results with control peptide (random sequence with TAT) treated animals. We anticipate that the findings from this study will have extensive clinical relevance.