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: 10231796
• 负责人: Doris Kretzschmar
• 金额: $ 19.53万
• 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/10231796
• 关键词: 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 (Brain); 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; Plant Roots; Population; Proteins; Reactive Oxygen Species; Resolution; Response Elements; Rodent Model; Role; Serotonin; Signal Transduction; Sleep; Slice; Statistical Methods; Steroids; Symptoms; Synapses; Testing; Triterpenes; Water; age related; age related neurodegeneration; base; design; experimental study; fly; gamma-Aminobutyric Acid; improved; in vivo; in vivo Model; inhibitor/antagonist; insight; madecassic acid; mating behavior; mouse model; neurotropic; relating to nervous system; 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.

项目摘要_项目 2 涉及植物膳食补充剂的有效临床试验的设计需要优化试验 产品中含有足够水平的活性化合物。这些活性物质的识别 因此，化合物必须成为任何认真的植物学研究的一部分。人们还认识到，总体 植物的活性特征可能是由多种活性化合物介导的，这些化合物可以独立发挥作用， 彼此相加、协同或拮抗。虽然植物的功能作用可以 在体内证明，活性化合物的鉴定及其相互作用的初步研究是最重要的 使用体外、离体或中等通量体内方法进行有效评估。这些也有 基于机制的优势。积雪草 (CA) 和睡茄 (WS) 已被证明 影响认知、睡眠和情绪，在这个项目中，我们将使用初级神经元分析、脑切片和 果蝇模型探索 CA 和 WS 的机制和活性化合物。初级神经元将是 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 树枝化、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和血管收缩弹性的影响，以测试是否 这些提取物促进血管健康。果蝇模型将用于识别可改善 与年龄相关的运动和反应能力下降。我们还将测试对所描述的类似抑郁状态的影响 在果蝇中，使用求偶作为一种分析方法，我们将测量神经元亚群中的血清素水平 被认为可以调节这种类似抑郁的状态。此外，果蝇将用于确定对 果蝇和哺乳动物的睡眠模式也会因年龄而改变。细胞和分子途径 调节睡眠在果蝇中是众所周知的，因此我们也可以研究 CA 和/或 WS 是否 通过改变神经元活动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。除了血清素和乙酰胆碱之外，还包括 GABA。到 将这些研究扩展到小鼠模型，我们将测量小鼠大脑切片中的神经元活动并解决 CA 和/或 WS 治疗是否会影响 GABA 信号传导（使用 GABA 抑制剂）。这些研究共同 将识别 CA 和 WS 中的活性化合物，并通过以下方式表明它们是否支持健康和恢复力： 促进认知、睡眠和/或情绪。此外，他们还将提供对可能的机制的见解 介导这些作用，例如改善神经元活动、减少氧化应激和/或促进 线粒体功能。未来的研究可以在体内小鼠模型中证实这些影响，并最终 为在人体中测试活性化合物提供基础。