Suppression of Immune Signatures of Stress by Polyphenols Supplements

摘要/摘要 项目1的目标是建立一种植物补充剂， 导致抑郁行为的病理生理反应。该提案将调查 生物活性膳食多酚制剂（BDPP）多酚代谢物如何与生物分子 小胶质细胞、中型多刺神经元（MSN）和血脑屏障（BBB）细胞中的遗传靶点，以促进 神经元功能和行为对压力反应的弹性。最近的研究表明， 免疫活性通过影响上述细胞的活性而赋予对应激的易感性。系统性 在模型中观察到应激引起的炎性细胞因子，特别是白细胞介素-6（IL-6）的上调 抑郁症患者以及MDD患者。从涉及验证模型的初步研究 我们发现，白细胞持续表达IL-6仅在应激状态下影响血脑屏障功能， 易感小鼠我们提出的证据表明，BDPP生物可利用的代谢物可能会促进社会弹性， 通过抑制细胞因子的产生来应激，这与维持BBB完整性有关。我们的第一 因此，我们的目标是确定BDPP是否通过靶向 与BBB功能相关的机制。这一目标将决定植物补充剂是否能预防 血脑屏障的通透性和外周免疫因子通过神经内皮的浸润，并将 表征内皮细胞中BDPP代谢物的生物分子靶标。我们的初步研究显示 补充BDPP可抑制应激诱导的小胶质细胞过度分化和 与压力易感性相关的炎症基因。我们的第二个目标是进行一次 无偏筛选，以鉴定小胶质细胞中响应应激的BDPP代谢物的分子生物活性。 这一目标涉及一种称为翻译核糖体亲和纯化的细胞特异性RNA测序技术 （陷阱）。此外，Aim 2将描述小胶质细胞中Toll/Toll样受体活性的增加如何赋予 对应激诱导的抑郁症的易感性，并作为BDPP代谢物的接近的生物靶标。 最后，我们展示了BDPP代谢物如何防止NAc中适应不良的神经元突触的产生， 通过调节MSN中突触基因的表达来响应压力。我们还发现外周血IL-6 外周血白细胞对应激反应的产生对于促进异常突触的形成是至关重要的， 的NAC。因此，最终目标将是进行无偏筛选，以确定 使用TRAP方法检测D2 MSN中的BDPP代谢物，并研究外周IL-6是否扩散到D2 MSN中。 脑通过IL-6 R激活小胶质细胞以实现NAc突触密度的调节。这将提供 为什么植物补充剂抑制外周IL-6可以促进弹性行为的机制 进行临床试验总之，项目1将验证与以下因素相关的病理生理反应： IL-6可以通过植物补充剂来抑制，以促进对压力的恢复力。

Summary/Abstract The goal of Project 1 is to establish that a botanical supplement provides resilience against stress-induced pathophysiological responses that confer susceptibility to depressive behavior. This proposal will investigate how polyphenol metabolites of a bioactive dietary polyphenol preparation (BDPP) engage biomolecular and genetic targets in microglia, medium spiny neurons (MSNs), and blood brain barrier (BBB) cells to promote resilience of neuronal function and behavior in response to stress. Recent studies show that dysfunctional immune activity confers susceptibility to stress by affecting activity of these aforementioned cells. Systemic upregulation of inflammatory cytokines, in particular interleukin-6 (IL-6), by stress is observed in model systems of depression, as well as in patients with MDD. From preliminary studies involving a validated model of social stress, we show persistent IL-6 expression from leukocytes affects the BBB function only in stress- susceptible mice. We present evidence that BDPP bioavailable metabolites may promote resilience to social stress by suppressing production of cytokines, which is associated with maintenance of BBB integrity. Our first goal is therefore to establish if BDPP provides resilience against stress-induced depression by targeting mechanisms associated with BBB function. This aim will determine whether a botanical supplement prevents BBB permeability and infiltration of peripheral immune factors through the neuroendothelium, and will characterize biomolecular targets of BDPP metabolites in endothelial cells. Our preliminary studies show BDPP supplementation suppresses stress-induced microglia hyper-ramification and upregulation of inflammatory genes, which are associated with stress-susceptibility. Our second goal is to conduct an unbiased screen to identify molecular bioactivities of BDPP metabolites in microglia in response to stress. This aim involves a cell-specific RNA-sequencing technique termed translating ribosomal affinity purification (TRAP). Moreover, Aim 2 will characterize how increased activity of toll/nod-like receptors in microglia confer susceptibility to stress-induced depression, and act as proximate biological targets for BDPP metabolites. Finally, we show how BDPP metabolites prevent maladaptive glutamatergic synapse generation in the NAc in response to stress by regulating expression of synaptic genes in MSNs. We also show that peripheral IL-6 production by peripheral leukocytes in response to stress is critical for facilitating aberrant synapse formation in the NAc. The final goal will therefore be to conduct an unbiased screen to identify molecular bioactivities of BDPP metabolites in D2 MSNs using the TRAP method and to investigate if peripheral IL-6 diffusion into the brain activates microglia via the IL-6R to effect regulation of NAc synapse densities. This would provide a mechanism for why suppression of peripheral IL-6 by a botanical supplement can promote resilient behaviors for a future clinical trial. Together, Project 1 will validate that pathophysiological responses associated with IL-6 can be suppressed by a botanical supplement to promote resilience to stress.