Department of Pharmacognosy, Faculty of Pharmacy, Hacettepe University, Ankara 06100, Turkey.
Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
Phytomedicine. 2020 Aug;74:152920. doi: 10.1016/j.phymed.2019.152920. Epub 2019 Apr 4.
Identification of the low abundance of phytochemicals in plant extracts is very difficult. Pharmacological activity observed in such plants is not due to a single compound. In most cases, plant extracts show activity based on synergistic or antagonistic effects. Therefore, the idea of a holistic approach is more rational.
This study was planned to compare the metabolomics and proteomics profiles of Valeriana officinalis L. (Valerianaceae), Melissa officinalis L. (Lamiaceae), Hypericum perforatum L. (Hypericaceae) and Passiflora incarnata L. (Passifloraceae) used in sedative anxiolytic and sleep disorders. Integrated omics analyses were used to provide a better understanding of the effect of plant extracts on the brain-derived neurotrophic factor (BDNF) expression levels on the SH-SY5Y cell line by a holistic approach.
Metabolomic profiling of the plants was performed using the GC-MS and LC-qTOF-MS systems, and the proteomics analysis using the LC-qTOF-MS system after trypsin digestion. The Human BDNF Quantikine ELISA kit was utilized to test BDNF expression activity on the SH-SY5Y cell line.
The investigated plant extracts showed a significant increase in BDNF expression (p < 0.05). M. officinalis was found as the most active extract. According to the correlation analyses between BDNF activity and metabolomics or proteomics level, 94 metabolites had a positive correlation while 23 metabolites had a highly negative correlation; those for proteins are 24 and 6, respectively.
The multivariate data analysis revealed a similar metabolomics profile of H. perforatum and P. incarnata, which also had a similar activity profile. Remarkably, all the primary metabolites belonging to the Krebs Cycle (citric acid, fumaric acid, succinic acid, pyruvic acid, malic acid and citramalic acid, an analog of malic acid) were positively correlated with BDNF activity. Secondary metabolites with a high BDNF expression belonged to flavonoids, xanthone, coumarines, tannin, naphtalenes, terpenoids and carotenoid skeleton. Two proteins from the cytochrome P450 family (P450 71B11 and P450 94B3) were positively correlated with BDNF activity. Employing omics technologies in the plant research area will offer a better understanding of the role of plant extracts and may lead to the discovery of new compounds with specific activity.
鉴定植物提取物中的低丰度植物化学物质非常困难。在这些植物中观察到的药理活性不是由单一化合物引起的。在大多数情况下，植物提取物基于协同或拮抗作用显示出活性。因此，整体方法的理念更为合理。
本研究旨在比较缬草（缬草科）、薄荷（唇形科）、贯叶连翘（金丝桃科）和西番莲（西番莲科）在镇静、抗焦虑和睡眠障碍中的代谢组学和蛋白质组学特征。综合组学分析用于通过整体方法提供对植物提取物对脑源性神经营养因子（BDNF）表达水平对 SH-SY5Y 细胞系影响的更好理解。
使用 GC-MS 和 LC-qTOF-MS 系统进行植物代谢组学分析，使用 LC-qTOF-MS 系统在胰蛋白酶消化后进行蛋白质组学分析。利用 Human BDNF Quantikine ELISA 试剂盒检测 SH-SY5Y 细胞系上 BDNF 表达活性。
研究发现，所研究的植物提取物均显著增加 BDNF 表达（p<0.05）。薄荷被发现是最活跃的提取物。根据 BDNF 活性与代谢组学或蛋白质组学水平之间的相关分析，有 94 种代谢物呈正相关，有 23 种代谢物呈高度负相关；蛋白质分别为 24 种和 6 种。
多元数据分析揭示了贯叶连翘和西番莲具有相似的代谢组学特征，也具有相似的活性特征。值得注意的是，所有属于三羧酸循环（柠檬酸、富马酸、琥珀酸、丙酮酸、苹果酸和苹果酸类似物）的主要代谢物均与 BDNF 活性呈正相关。具有高 BDNF 表达的次生代谢物属于黄酮类、黄烷酮、香豆素、单宁、萘、萜类和类胡萝卜素骨架。细胞色素 P450 家族的两种蛋白（P450 71B11 和 P450 94B3）与 BDNF 活性呈正相关。在植物研究领域中采用组学技术将提供对植物提取物作用的更好理解，并可能导致发现具有特定活性的新化合物。
