四物汤对辐射致血虚证小鼠作用的代谢组学研究

Metabonomic study on Siwu Tang in radiation-induced blood deficient mice

目的:基于超高效液相色谱-四级杆飞行时间质谱联用(UPLC-ESI-Q-TOF-MS)技术进行辐射损伤致血虚证以及四物汤的方证结合研究,发现辐射致血虚证及用四物汤干预后小鼠内源性代谢物的变化,基于代谢组学的方法考察血虚证的发生及四物汤的作用机制。方法:选用C57小鼠36只,按体重随机分为对照组、模型组及四物汤组。造模方法采用3.5Gy60Coγ射线全身照射制作模型,造模7 d后股动脉取血并摘取脾脏和胸腺,进行处理后用UPLC-ESI-Q-TOF-MS检测,应用Mark-erLynx XS软件对总离子流图进行色谱峰识别,利用主成分分析法对数据进行处理,采用正交偏最小二乘法进行组间数据分析。通过模式识别分析,快速确认标志物。结果:辐射损伤所致的血虚证小鼠血清中溶血磷脂、葡糖苷酸、单酰甘油酯、三羟基丁酸、神经酰胺、天冬氨酸磷酸酯、甘油磷脂等代谢物的含量变化明显;脾脏中变化明显的代谢物有单酰甘油酯、神经酰胺、溶血磷脂、羟基丁酸、棕榈酸、3-羟基硬脂酸、二甲基精氨酸、苯丙氨酸、磷脂酰丝氨酸;胸腺中有甲基腺苷、谷甾醇、肉毒碱、磷脂酰肌醇、磷脂酰乙醇胺、甘油二酯、二甲基精氨酸、神经酰胺、溶血磷脂、羟基丁酸、胆固醇等。结论:通过对这些生物标志物的生理功能分析,推测辐射损伤使小鼠机体的脂类代谢、糖类代谢及部分氨基酸的合成代谢发生紊乱,经典补血方剂四物汤治疗可以使之逆转并调整平衡。

Objective： To study the radiation-induced blood deficiency and the combination of prescription and syndrome of Siwu Tang using ultra performance liquid chromatography-quadrupoles-time of flight mass spectrometer（UPLC-ESI-Q-TOF-MS）,in order to discover the changes in metabolic profiles of blood deficient mice induced by radiation,and clarify the relationship between blood deficient syndrome and the mechanism of Siwu Tang.Method： Thirty six C57 mice were randomly divided into three groups： the control group and the model model groups and the Siwu Tang group.The model was established by general irradiation with 3.5Gy 60Coγ ray.The animals were sacrificed on the 7th day after radiation and their blood,spleens and thymus were collected and detected using UPLC-ESI-Q-TOF/MS.MarkerLynx XS software was adopted to identify chromatographic peaks in the total ion chromatogram.Data was processed by making principal component analysis and analyzed by orthogonal partial least squares method among these groups,in order to rapidly identify marks through pattern recognition and analysis.Result： Compared with the control group,lysophosphatide,glucosiduronic acid,monoacylglycerol,erythronic acid,ceramide,aspartate phosphate ester,glyceryl phosphatide were obviously changed in the sera of the model group.Monoacylglycerol,ceramide,lysophosphatide,hydroxybutyric acid,palmitinic acid,3-hydroxy-stearic acid,diethylarginine,neuraminic acid,phosphatidylserine were found in metabolites of their spleens.Methyladenosine,sitosterol,carnitine,phosphatidylinositol,phosphatidylethanolamine,diglyceride,dimethylarginine,ceramide,hydroxybutyric acid,cholesterol were found in thymus of the model group.Conclusion： Analysis on physiological functions of these biological markers show that radiation could induce the disorder of the metabolism of lipoid and carbohydrate and affect the synthesis of some amino acids,and Siwu Tang can reverse these effects.