综合放血法所致血虚证小鼠模型及四物汤反证的代谢组学研究  

NMR-spectroscopy-based metabonomic study on mouse model of blood deficiency syndrome induced by compound method of bleeding,starved feeding and exhausting and effect of Siwutang

作　　者：霍超[1] 马增春[1] 王穆[1] 刘娟[2] 颜贤忠[2] 高月[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所药理与毒理研究室,北京100850 [2]军事医学科学院生物医学分析中心,北京100850

出　　处：《中国中药杂志》2010年第23期3194-3198,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(30672628;30730112)

摘　　要：目的:通过检测综合放血法所致血虚证及用四物汤干预治疗后小鼠内源性代谢物的变化,确定综合放血法所致血虚证小鼠的血清及组织相关的生物标记物,从代谢组学角度研究血虚证证本质及四物汤的作用机制。方法:C57小鼠30只,按体重随机分为3组:正常对照组、综合放血模型组、四物汤反证组,每组10只。造模10 d后于小鼠股动脉取血,同时取脾脏、胸腺及股骨骨髓,并分别提取血清及组织的脂溶性和水溶性代谢物。用600 MHz超导傅立叶变换核磁共振波谱仪检测,对所得到的氢谱按每段0.01进行分段积分并归一化,用SIMCA-P10.0软件进行主成分分析。结果:与对照组相比,模型组的血清及各组织的内源性代谢物中乳酸、胆碱、牛磺酸、丙氨酸、低密度脂肪酸、甘油、不饱和脂肪酸、肌酐、乙酰乙酸盐、丙氨酸及β-羟基丁酸的含量有明显变化,这些变化在四物汤治疗组中有不同程度的回调。上述代谢物可以作为潜在的生物标记物进行相关机制的研究。结论:综合放血法制作的血虚证小鼠的糖有氧氧化受到抑制,无氧酵解减弱,脂肪动员加强,淋巴细胞增殖受到抑制及机体免疫功能受损。四物汤能够对上述血虚证病理结果进行逆转。Objective： To explore the essence of blood deficiency syndrome and the mechanism of the effects of Siwutang based on the characteristic changes in metabolic profiles of C57 mice with blood deficiency induced by compound method of bleeding,starved feeding and exhausting before and after the treatment of Siwutang.Method： Thirty C57 mice were randomly divided into three groups of 10 animals： the control group and two blood deficiency model groups without or with Siwutang treatment.The animals were sacrificed after induction by compound method of bleeding,starved feeding and exhausting for 10 days and the blood,spleen,thymus and bone marrow were obtained.The metabolic profiles of serum,aqueous and lipidic extracts of thymus,spleen and bone marrow were investigated using 1H nuclear magnetic resonance spectroscopy（NMR）.The NMR spectra were integrated in segments of 0.01,normalized and then analyzed using SIMCA-P software to visualize the similarities and differences in metabolic profiles between these groups.Result： Compared with the control group,the model group contained different levels of lactate,choline,glucose,taurine,alanine,LDL,glycerin,UFA,creatinine,acetoacetate,glutamate and β-hydroxybutyrate（β-HB）.In the Siwutang treated group,the above-mentioned changes were reversed.Conclusion： The disorder of energy metabolism,damage of lymphocyte,and disorder of immune function were observed in blood deficiency model by NMR-based-metabonomics method,and the Siwutang can improve these effects.Metabonomics may be a valuable tool in the pharmacological evaluation of Siwutang.

关 键 词：四物汤 血虚证 代谢组学 核磁共振 主成分分析 

分 类 号：R285[医药卫生—中药学]