偏头痛大鼠的血浆代谢组学研究

Plasma metabonomic analysis of migraine in rats

目的运用代谢组学方法,研究大鼠偏头痛的发病机制。方法采用改进的Christina tassorelli方法建立大鼠偏头痛模型(模型组)并设正常对照组。取大鼠血样,通过超高效液相色谱仪结合高分辨率质谱仪联用(UPLCQ-TOF/MS)对模型组与正常对照组血浆进行代谢谱检测。通过Micromass Markerlynx软件对采样数据进行归一化处理。统计学分析使用SIMCA-P+软件偏最小二乘法进行模式识别。结果 UPLC-Q-TOF代谢色谱图显示,与正常对照组相比,模型组的UPLC图出峰量明显增多,而且两组图谱之间有一定程度的关联性。散点分布图显示,正常对照组和模型组血样标本均具有明显的聚类作用,且有趋势显著的分类运动。模型组与正常对照组的荷载图并结合MS-MS的串联质谱结果显示,5-羟吲哚乙酸、3,4-二羟苯乙二醇、磷脂酰胆碱为潜在的生物标志物。结论本研究建立了基于大鼠血样的UPLC-Q-TOF/MS代谢组学研究方法,初步阐释了偏头痛发病的生物标志物及代谢组学机制。

Objective To investigate the pathogenesis of rat migraine using the metabonomic method. Methods The mi- graine model of Wistar rat was established with the improved Christina tassorelli method （ model group） , and normal control group was set up. Taking rat blood samples, the plasma metabolic profiling in two groups was detected by the uhraperfor- mance liquid chromatograph （ UPLC ） combined with high resolution mass spectrometer （ Q-TOF/MS） （ UPLC-Q-TOF / MS）. The sample data were normalized by the Micromass Markerlynx software, and then the pattern recognition of data was made by partial least square discrinfinant analysis （ PLS-DA ） of SIMCA-P ＋ software. Results UPLC-Q-TOF metabolic chromatogram showed that the peak number of UPLC ehromatogram in model group increased significantly compared with normal control group, and there was a relation of certain degree between the chromatograms of two groups. Scatter diagram showed that the blood samples in both two groups had significant clustering effect and a trend of significant classification movement. The results of analysis of loading plot combined with tandem mass spectrometry （MS-MS） showed that 5-hydrox- yindoleacetic acid,3,4-dihydroxyphenyl glycol and phosphatidyl choline were the potential biomarkers. ConcLusion This study established a UPLC-Q-TOF/MS metabonomic research method based on the rat blood samples and preliminary interpreted the biomarkers and the metabonomic mechanism of migraine pathogenesis.