Mechanism based evaluation of botanical bioactive compounds enhancing neurological resilience (Project 2)

基于机制的植物生物活性化合物增强神经恢复能力的评估（项目2）

• 批准号: 10661729
• 负责人: Doris Kretzschmar
• 金额: $ 15.73万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661729
• 关键词: Acetylcholine; Acids; Address; Affect; Age; Aging; Antioxidants; Arylalkylamine N-Acetyltransferase; Ashwagandha; Attention; Ayurvedic Medicine; Biological Assay; Blood Vessels; Botanical dietary supplements; Botanicals; Brain; Chemicals; Choline O-Acetyltransferase; Clinical Trials; Cognition; Cognitive; Coupled; Courtship; Crude Extracts; Dose; Drosophila genus; Drosophila inturned protein; Drosophila melanogaster; Elderly; Electron Transport; Evaluation; Fingerprint; Future; GABA Receptor; Gene Expression; Glycosides; Goals; Gotu kola; Head; Health; Herb; Hippocampus; Human; Immunohistochemistry; In Vitro; Individual; Invertebrates; Lactones; Liquid Chromatography; Locomotion; Mammals; Mass Spectrum Analysis; Materials Testing; Measures; Mediating; Mental Depression; Mitochondria; Modeling; Molecular; Moods; Mus; Neurologic; Neurons; Neurotransmitters; Oxidative Stress; Parents; Pathway interactions; Population; Proteins; Reactive Oxygen Species; Resolution; Response Elements; Rodent Model; Role; Serotonin; Signal Transduction; Sleep; Slice; Statistical Methods; Symptoms; Synapses; Testing; Triterpenes; Water; age related; age related neurodegeneration; constriction; design; experimental study; fly; gamma-Aminobutyric Acid; improved; in vivo; in vivo Model; inhibitor; insight; madecassic acid; mating behavior; mouse model; neural; neurotropic; promote resilience; resilience; response; sleep pattern; sleep regulation; vasoconstriction; vibration

PROJECT SUMMARY_PROJECT 2 The design of valid clinical trials involving botanical dietary supplements requires the optimization of the trial product such that it contains sufficient levels of the active compounds. The identification of these active compounds therefore has to be a part of any serious study of botanicals. It is also recognized that the overall activity profile of a botanical may be mediated by multiple active compounds, which can act independently, additively, synergistically or be antagonistic to each other. While functional effects of a botanical can be demonstrated in vivo, initial studies on the identification of active compounds and their interaction is most efficiently evaluated using in vitro, ex vivo, or moderate throughput in vivo approaches. These also have the advantage of being mechanism based. Centella asiatica (CA) and Withania somnifera (WS) have been shown to affect cognition, sleep and mood and in this project, we will use primary neuron assays, brain slices, and Drosophila models to explore mechanisms and active compounds of CA and WS. Primary neurons will be used to address effects of CA and WS and their compounds on neuronal health by measuring neuronal arborization, antioxidant responses, reactive oxygen species, and mitochondrial function. In addition, we will determine effects on vascular tone and resilience to vasoconstriction in mouse brain slices to test whether these extracts promote vascular health. Drosophila models will be used to identify compounds that improve the age-related decline in locomotion and reactivity. We will also test effects on a described depression-like state in Drosophila, using courtship as an assay, and we will measure Serotonin levels in neuronal subpopulations identified as mediating this depression-like state. Furthermore, Drosophila will be used to determine effects on sleep patterns, which are also altered by age in flies and mammals. The cellular and molecular pathways regulating sleep are well known in Drosophila and we can therefore also investigate whether CA and/or WS promotes healthy sleep patterns by altering neuronal activity and neurotransmitter signaling in the neuronal populations that promote or suppress sleep. Besides Serotonin and Aceteylcholine, this includes GABA. To expand these studies to the mouse model, we will measure neuronal activity in mouse brain slices and address whether treatment with CA and/or WS affects GABA signaling (using GABA inhibitors). Together, these studies will identify active compounds in CA and WS and they will show whether they support health and resilience by promoting cognition, sleep and/or mood. In addition, they will provide insights into mechanism that may mediate these effects, like improving neuronal activity, decreasing oxidative stress and/or promoting mitochondrial function. Future studies can then confirm these effects in an in vivo mouse model and eventually provide the basis for testing active compounds in humans.

项目summary_project 2 涉及植物饮食补充剂的有效临床试验的设计需要优化试验 产品使其包含足够水平的活性化合物。这些活动的识别 因此，化合物必须成为对植物药的任何认真研究的一部分。还认识到总体 植物学的活性曲线可能是由多种活性化合物介导的，这些化合物可以独立起作用， 添加，协同或彼此对抗。虽然植物的功能效应可能是 在体内证明，有关活性化合物鉴定及其相互作用的初步研究最多是 使用体外，体内或中等吞吐量在体内方法进行有效评估。这些也有 基于机制的优势。已经显示了Centella Asiatica（CA）和Withania Somnifera（WS） 为了影响认知，睡眠和情绪，在这个项目中，我们将使用主要的神经元测定，脑切片和 果蝇模型探索CA和WS的机制和活性化合物。主要神经元将 用于通过测量神经元来解决CA和WS及其化合物对神经元健康的影响 树木化，抗氧化反应，活性氧和线粒体功能。此外，我们将 确定对小鼠脑切片中血管张力的影响和对血管收缩的弹性，以测试是否 这些提取物促进了血管健康。果蝇模型将用于识别改进的化合物 与年龄相关的运动和反应性下降。我们还将测试对所描述的抑郁状态的影响 在果蝇中，使用求爱作为测定法，我们将测量神经元亚群中的5-羟色胺水平 被确定为介导这种抑郁状态。此外，果蝇将用于确定对 睡眠模式也随着苍蝇和哺乳动物的年龄而改变。细胞和分子途径 调节睡眠在果蝇中是众所周知的，因此我们还可以研究CA和/或WS是否 通过改变神经元的神经元活性和神经递质信号传导来促进健康的睡眠模式 促进或抑制睡眠的人群。除了5-羟色胺和乙酰胆碱外，还包括GABA。到 将这些研究扩展到小鼠模型，我们将测量小鼠脑切片中的神经元活性并解决 使用CA和/或WS处理是否影响GABA信号传导（使用GABA抑制剂）。在一起，这些研究 将在CA和WS中识别活性化合物，它们将表明它们是否支持健康和韧性 促进认知，睡眠和/或情绪。此外，他们将提供有关可能的机制的见解 介导这些作用，例如改善神经元活性，减少氧化应激和/或促进 线粒体功能。然后，未来的研究可以在体内小鼠模型中确认这些效果，并最终确认 提供了测试人类活性化合物的基础。