辐射损伤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 the mouse model of blood deficiency syndrome induced by radiation

[目的]通过检测辐射损伤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,确定辐射损伤所致血虚证小鼠的血清及组织相关的生物标记物,基于代谢组学的方法考察血虚证证本质及四物汤的作用机制。[方法]C57小鼠按体质量随机分为正常组、辐射造模组、四物汤反证组。造模7d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。[结果]主成分分析结果显示正常组、血虚证模型组以及四物汤反证组能够明显的区分。[结论]四物汤治疗血虚证的机制可能与稳定细胞膜,维持细胞内外渗透压平衡;调节糖酵解,促进机体正常能量代谢以及增强机体抗氧化能力、免疫力有关。

[Objection] 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 radiation before the treatment of Siwutang.[Methods] 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 the 7th day after radiation 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 ppm,normalized and then analyzed using SIMCA-P software to visualize the similarities and differences in metabolic profiles between these groups.[Results] Compared with the control group,the model group contained different levels of lactate,choline,glucose,taurine,alanine,LDL,glycerin,UFA,creatinine,histidine,valine/leucine/isoleucine.In the Siwutang treated group,the above-mentioned changes were reversed.[Conclusion] The injury established by radiation was a kind of proper model to develop further metabonomics researches.The disorder of energy metabolism and osmoregulation,and the damage of immune function were observed by NMR-based-metabonomics method.Siwutang,the classic complex for treating blood deficiency in clinic,could improve the immune function,accommodate energy metabolism and stabilize cellular membrane.