Zhou Hui, Qu Zhe, Mossine Valeri V, Nknolise Dineo L, Li Jilong, Chen Zhenzhou, Cheng Jianlin, Greenlief C Michael, Mawhinney Thomas P, Brown Paula N, Fritsche Kevin L, Hannink Mark, Lubahn Dennis B, Sun Grace Y, Gu Zezong
Department of Pathology and Anatomical Sciences, University of Missouri School of Medicine, Columbia, Missouri, United States of America; Center for Translational Neuroscience, University of Missouri School of Medicine, Columbia, Missouri, United States of America.
Department of Biochemistry, University of Missouri School of Medicine, Columbia, Missouri, United States of America.
PLoS One. 2014 Nov 24;9(11):e113531. doi: 10.1371/journal.pone.0113531. eCollection 2014.
Aged garlic extract (AGE) is widely used as a dietary supplement, and is claimed to promote human health through anti-oxidant/anti-inflammatory activities with hypolipidemic, antiplatelet and neuroprotective effects. Prior studies of AGE have mainly focused on its organosulfur compounds, with little attention paid to its carbohydrate derivatives, such as N-α-(1-deoxy-D-fructos-1-yl)-L-arginine (FruArg). The goal of this study is to investigate actions of AGE and FruArg on antioxidative and neuroinflammatory responses in lipopolysaccharide (LPS)-activated murine BV-2 microglial cells using a proteomic approach. Our data show that both AGE and FruArg can significantly inhibit LPS-induced nitric oxide (NO) production in BV-2 cells. Quantitative proteomic analysis by combining two dimensional differential in-gel electrophoresis (2D-DIGE) with mass spectrometry revealed that expressions of 26 proteins were significantly altered upon LPS exposure, while levels of 20 and 21 proteins exhibited significant changes in response to AGE and FruArg treatments, respectively, in LPS-stimulated BV-2 cells. Notably, approximate 78% of the proteins responding to AGE and FruArg treatments are in common, suggesting that FruArg is a major active component of AGE. MULTICOM-PDCN and Ingenuity Pathway Analyses indicate that the proteins differentially affected by treatment with AGE and FruArg are involved in inflammatory responses and the Nrf2-mediated oxidative stress response. Collectively, these results suggest that AGE and FruArg attenuate neuroinflammatory responses and promote resilience in LPS-activated BV-2 cells by suppressing NO production and by regulating expression of multiple protein targets associated with oxidative stress.
aged大蒜提取物（AGE）作为一种膳食补充剂被广泛使用，据称它通过抗氧化/抗炎活性具有降血脂、抗血小板和神经保护作用，从而促进人类健康。先前对AGE的研究主要集中在其有机硫化合物上，而对其碳水化合物衍生物，如N-α-（1-脱氧-D-果糖-1-基）-L-精氨酸（FruArg）关注较少。本研究的目的是使用蛋白质组学方法研究AGE和FruArg对脂多糖（LPS）激活的小鼠BV-2小胶质细胞中抗氧化和神经炎症反应的作用。我们的数据表明，AGE和FruArg均可显著抑制LPS诱导的BV-2细胞中一氧化氮（NO）的产生。通过二维差异凝胶电泳（2D-DIGE）与质谱联用的定量蛋白质组学分析表明，LPS暴露后26种蛋白质的表达发生了显著变化，而在LPS刺激的BV-2细胞中，分别有20种和21种蛋白质的水平在AGE和FruArg处理后表现出显著变化。值得注意的是，对AGE和FruArg处理有反应的蛋白质中约78%是相同的，这表明FruArg是AGE中的主要活性成分。MULTICOM-PDCN和 Ingenuity Pathway分析表明受AGE和FruArg处理差异影响的蛋白质参与炎症反应和Nrf2介导的氧化应激反应。总的来说，这些结果表明AGE和FruArg通过抑制NO的产生以及调节与氧化应激相关的多个蛋白质靶点的表达，减轻LPS激活的BV-2细胞中的神经炎症反应并促进其恢复能力。
