Seo Han-Seok, Hirano Misato, Shibato Junko, Rakwal Randeep, Hwang In Kyeong, Masuo Yoshinori
Department of Food and Nutrition, College of Human Ecology, Seoul National University, Seoul 151-742, South Korea.
J Agric Food Chem. 2008 Jun 25;56(12):4665-73. doi: 10.1021/jf8001137. Epub 2008 Jun 3.
The aim of this study was 2-fold: (i) to demonstrate influences of roasted coffee bean aroma on rat brain functions by using the transcriptomics and proteomics approaches and (ii) to evaluate the impact of roasted coffee bean aroma on stress induced by sleep deprivation. The aroma of the roasted coffee beans was administered to four groups of adult male Wistar rats: 1, control group; 2, 24 h sleep deprivation-induced stress group (the stress group); 3, coffee aroma-exposed group without stress (the coffee group); and 4, the stress with coffee aroma group (the stress with coffee group). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of some known genes responsive to aroma or stress was performed using total RNA from these four groups. A total of 17 selected genes of the coffee were differently expressed over the control. Additionally, the expression levels of 13 genes were different between the stress group and the stress with coffee group: Up-regulation was found for 11 genes, and down-regulation was seen for two genes in the stress with coffee group. We also looked to changes in protein profiles in these four samples using two-dimensional (2D) gel electrophoresis; 25 differently expressed gel spots were detected on 2D gels stained by silver nitrate. Out of these, a total of nine proteins were identified by mass spectrometry. Identified proteins belonged to five functional categories: antioxidant; protein fate; cell rescue, defense, and virulence; cellular communication/signal transduction mechanism; and energy metabolism. Among the differentially expressed genes and proteins between the stress and the stress with coffee group, NGFR, trkC, GIR, thiol-specific antioxidant protein, and heat shock 70 kDa protein 5 are known to have antioxidant or antistress functions. In conclusion, the roasted coffee bean aroma changes the mRNA and protein expression levels of the rat brain, providing for the first time clues to the potential antioxidant or stress relaxation activities of the coffee bean aroma.
（i）通过转录组学和蛋白质组学方法证明烘焙咖啡豆香气对大鼠脑功能的影响；（ii）评估烘焙咖啡豆香气对睡眠剥夺诱导的应激的影响。将烘焙咖啡豆的香气施加于四组成年雄性Wistar大鼠：1，对照组；2，24小时睡眠剥夺诱导应激组（应激组）；3，无应激的咖啡香气暴露组（咖啡组）；4，应激与咖啡香气组（应激与咖啡组）。使用这四组的总RNA对一些已知的对香气或应激有反应的基因进行逆转录聚合酶链反应（RT-PCR）分析。与对照组相比，共17个选定的咖啡基因表达不同。此外，应激组和应激与咖啡组之间13个基因的表达水平不同：应激与咖啡组中有11个基因上调，2个基因下调。我们还使用二维（2D）凝胶电泳观察了这四个样品中蛋白质谱的变化；在硝酸银染色的2D凝胶上检测到25个表达不同的凝胶斑点。其中，通过质谱鉴定了总共9种蛋白质。鉴定出的蛋白质属于五个功能类别：抗氧化剂；蛋白质命运；细胞拯救、防御和毒力；细胞通讯/信号转导机制；以及能量代谢。在应激组和应激与咖啡组之间差异表达的基因和蛋白质中，已知神经生长因子受体（NGFR）、酪氨酸激酶C（trkC）、G蛋白偶联内向整流钾通道（GIR）、硫醇特异性抗氧化蛋白和热休克70 kDa蛋白5具有抗氧化或抗应激功能。总之，烘焙咖啡豆香气改变了大鼠脑的mRNA和蛋白质表达水平，首次为咖啡豆香气潜在的抗氧化或应激缓解活性提供了线索。
