Gao Shan-Shan, Guo Hui-Qing, Zhang Ze-Kun, Bai Guang-Can, Gao Xiao-Yan, Ma Chang-Hua
School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100102, China.
Zhongguo Zhong Yao Za Zhi. 2017 Jun;42(12):2373-2379. doi: 10.19540/j.cnki.cjcmm.20170313.001.
In order to reveal the properties of polar metabolome in inflammatory cells, we selected LPS-induced RAW264.7 inflammatory cell models as the carrier for the research of metabolic fingerprint analysis. In this study, an ultra performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS)-based metabolomics protocol was optimized for the extraction of polar metabolites from RAW264.7 cell line. Then orthogonal partial least squares discriminant analysis (OPLS-DA) was used to process the metabolic data, and finally, a total of 17 metabolites were selected and identified. The results showed that MeOH-CHCl3-H2O (8∶1∶1) was chosen as the optimal extraction solvent to achieve higher number of chromatographic peaks, with the best relative extraction efficiency and stability. Comparing with the normal cells, the inflammatory cells presented an abnormal metabolism in protein, carbohydrate, nucleotide and phospholipids. In this study, a UPLC-Q-TOF/MS-based metabolomics protocol for the polar metabolites from RAW264.7 cell line was developed, which may provide important information for the study of mechanism of inflammation and the anti-inflammatory drugs.
为了揭示炎性细胞中极性代谢组的特性，我们选择脂多糖诱导的RAW264.7炎性细胞模型作为代谢指纹分析研究的载体。在本研究中，优化了基于超高效液相色谱-四极杆飞行时间质谱联用仪（UPLC-Q-TOF/MS）的代谢组学方法，用于从RAW264.7细胞系中提取极性代谢物。然后采用正交偏最小二乘法判别分析（OPLS-DA）处理代谢数据，最终共筛选并鉴定出17种代谢物。结果表明，选择甲醇-氯仿-水（8∶1∶1）作为最佳提取溶剂可获得更多的色谱峰，具有最佳的相对提取效率和稳定性。与正常细胞相比，炎性细胞在蛋白质、碳水化合物、核苷酸和磷脂方面呈现代谢异常。本研究建立了一种基于UPLC-Q-TOF/MS的RAW264.7细胞系极性代谢物代谢组学方法，可为炎症机制研究及抗炎药物研究提供重要信息。
