Excellence in Research of dietary blueberry delays aging and reverses aging-induced physical dysfunctions by regulating PPAR pathway

这项研究旨在了解蓝莓的饮食摄入量如何发挥作用，特别是对65岁以上的人来说。蓝莓已被证明对健康有积极影响。该项目将利用小鼠，重点研究蓝莓在分子、细胞和组织水平上的抗衰老机制。如果初步结果在计划在老鼠身上进行的长期实验中成立，那么这项研究可能会导致人类通过食用天然蓝莓产品来促进健康和延长寿命。通过研究影响肌肉和血管的分子途径的新实验，将检验两个重要的想法：(1)饮食摄入蓝莓可以延长健康寿命，即使从高龄开始也是如此；(2)饮食摄入蓝莓可以延缓正常的衰老过程。除了了解某些类型的食物如何促进健康的明确社会效益外，该项目还将为历史悠久的黑人大学的少数民族学生(6名研究生和6名本科生)提供深入的研究培训。虽然衰老过程的确切机制尚不清楚，但延缓衰老引起的身体机能障碍和相关疾病的发展是促进健康老龄化的必要和有效的战略。PI最近报道，饮食中摄入表儿茶素(EC)，包括蓝莓在内的各种食物的生物活性成分，可以逆转衰老引起的骨骼肌(SKM)功能障碍，并提高老龄小鼠的存活率。兔眼蓝莓的EC含量是所有新鲜水果中最高的。该项目将检验这样一种假设，即饮食蓝莓EC通过调节PPAR/NF-kB通路来逆转衰老引起的SKM和血管内皮功能障碍。两个主要目标构成了实验工作的结构。目的1：以剂量依赖的寿命方案测试饮食蓝莓和EC的作用。将兔眼蓝莓冻干粉(高EC)、高丛蓝莓冻干粉(低EC)或纯EC冻干粉分别喂给20月龄C57BL/6小鼠(7组，每组30只，雌雄各30只)，直至自然生命结束。完整的Kaplan-Meier生存曲线和DNA甲基化时钟将用于监测衰老过程。目的：研究PPAR/NF-kB通路在骨骼肌和血管内皮细胞抗衰老中的作用。将兔眼蓝莓全粉，不加/不加PPAR特异性拮抗剂GW9662的纯EC喂给20月龄小鼠15周，同时进行跑台运动和体外试验。包括SKM、主动脉和血清在内的组织和体外培养的细胞将通过进行转录组和代谢组分析来确定PPAR/NF-kB通路的作用和其他抗衰老作用的机制。作为饮食对生化途径影响的实验研究，这项研究总体上将有助于推进这类人类健康研究的方法学，特别是建立蓝莓摄入延缓衰老引起的骨骼肌和血管系统功能障碍，进而延长人类寿命的机制。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This study aims to understand how the dietary intake of blueberries, which have been shown to have positive effects on health, might work to do so, particularly with regards to people over 65 years of age. Using mice, the project will focus on blueberry’s anti-aging mechanisms at the level of molecules, cells, and tissues. If the preliminary results hold true in the planned longer-term experiments on mice, then the research may lead to work in humans to promote health and extend lifespan by consuming natural blueberry products. With novel experiments examining molecular pathways that impact muscle and blood vessels, two important ideas will be tested: (1) dietary intake of blueberry can extend a healthy lifespan, even starting from advanced age, and (2) dietary intake of blueberry can delay the normal aging process. In addition to the clear societal benefits of understanding how some types of food work to promote health, this project will also provide in-depth research training to minority students (6 graduate students and 6 undergraduates) at a Historically Black University.While the exact mechanisms underlying the aging process are not well understood, delaying the aging-induced physical dysfunctions and related disease development is an essential and effective strategy to promote healthy aging. The PI recently reported that dietary intake of epicatechin (EC), a bioactive component of various foods, including blueberries, reversed aging-induced skeletal muscle (SkM) dysfunction and promoted the survival rate of aged mice. Rabbiteye blueberries contain the highest EC of all fresh fruits. The project will test the hypothesis that dietary blueberry EC reverses the aging-induced dysfunctions of SkM and vascular endothelium by regulating PPAR/NF-kB pathways in aged mice. Two primary objectives structure the experimental work. Objective 1: Test the effects of dietary blueberry and EC in a dose-dependent lifespan protocol. Freeze-dried powders of rabbiteye blueberry (high EC), highbush blueberry (low EC), or pure EC will be administrated to 20-month-old C57BL/6 mice (7 groups, 30 male, 30 female mice/group) until the natural end of lifespan. Complete Kaplan-Meier survival curves and DNA methylation clocks will be used to monitor the aging process. Objective 2: Test that PPAR/NF-kB pathways mediate blueberry EC anti-aging effect in skeletal muscle and vasculature. The whole powder of rabbiteye blueberry, pure EC with/out PPAR specific antagonist GW9662, will be fed to 20-month-old mice for 15 weeks while treadmill performance and ex vivo assays are conducted in parallel. Tissues including SkM, aorta, and serum and in vitro cultured cells will be used to determine the roles of PPAR/NF-kB pathways and other mechanisms of the anti-aging effect by conducting transcriptome and metabolome analyses. As an experimental study of the impact of diet on biochemical pathways, this study will help advance the methodologies for this type of human-health research, in general, and to establish, in particular, the mechanisms by which blueberry intake delays aging-induced dysfunction of skeletal muscle and vasculature and then extends the lifespan in humans.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.