四物汤对环磷酰胺所致血虚证治疗后的代谢组学研究  

NMR-spectroscopy-based metabonomic approach to analysis of Siwutang,a novel prescription,treated blood deficiency in mice

作　　者：王穆[1,2] 让蔚清[2] 张琪[3] 霍超[1] 马增春[1] 王宇光[1] 颜贤忠[3] 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]南华大学公共卫生学院环境医学与放射卫生研究所,湖南衡阳421001 [3]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5期630-634,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:通过分析血虚证模型以及给予四物汤反证后小鼠血清和组织提取物的代谢组学变化,结合模式识别的统计分析方法,探讨环磷酰胺所致血虚证及四物汤治疗后的特征性内源代谢谱的改变,为中药经典复方四物汤的作用机制研究提供科学依据。方法:将C57小鼠按体重随机分为3组:正常对照组、环磷酰胺模型组、四物汤给药组(10 g.kg-1.d-1),每组10只,连续给药7 d后股动脉取血,同时取脾脏、胸腺及股骨骨髓。用600 MHz超导傅立叶变化核磁共振波谱仪检测,对所得到的氢谱按每段0.04进行分段积分并归一化后,用SIMCA-P10.0软件进行主成分分析。结果:正常对照组、环磷酰胺模型组和四物汤给药组的血清代谢物组成、胸腺、脾脏、骨髓提取物的成分明显不同。其中环磷酰胺导致血清中的乳酸、3-羟基丁酸、胆碱、葡萄糖的含量下调,低密度脂蛋白、亮氨酸/异亮氨酸含量上调;胸腺提取物中的牛磺酸,胆碱,F:βRCH2CH2CO,epi-coprostanol,乳酸含量下调;脾脏提取物中的乳酸,3-羟基丁酸,胆碱,F:βRCH2CH2CO,epi-coprostanol,牛磺酸的含量下调;骨髓提取物中的乳酸、3-羟基丁酸、胆碱、葡萄糖的含量下调。给予四物汤后,以上各成分在不同程度上得到有效逆转。结论:环磷酰胺所致血虚证的可能机制是导致机体脂肪酸β氧化功能紊乱、细胞内外渗透压平衡破坏以及糖酵解功能失调。经典补血方剂治疗血虚证的机制可能与改善线粒体功能,纠正脂肪酸β氧化功能紊乱;稳定细胞膜,维持细胞内外正常渗透压;调节糖酵解,促进机体正常能量代谢有关。Objective： To discover the characteristic changes of metabolic profiles in C57 mice with cyclophosphamide induced blood deficiency and the effect of Siwutang. Method ： An integrated metabonomic study using high-resolution i H nuclear magnetic resonance （NMR） spectroscopy has been applied to investigate the metabolic profiles of serum, aqueous and lipidic extracts of thoracic gland, spleen, bone marrow obtained from control, model group （ intraperitoneal injection of cyclophosphamide at a dose of 250 mg · kg^- 1 ） and Siwutang treated model group. The NMR spectra were integrated in segments of 0. 04 ppm and then analyzed by principal component analysis （PCA） using SIMCA-P software to visualize the similarities and differences in metabolic profiles between these groups. Result： PCA result showed conspicuous difference in the metabolic profiles between groups. Compared with the control group, the model group contained lower concentration of lactate, 3-hydroxybutyrate, choline, glucose, and higher concentration of VLDL/LDL, leucine/isoleucine in serum. Lower concentration of taurine choline, Fβ： RCH2 CH2 CO, Epi-coprostanol and lactate were found in both in thoracic gland extracts and spleen extracts. And in spleen extracts, we also found the lower concentration of 3-HB. In the extracts of bone marrow, the lower concentration of lactate, choline, glueosee were observed, When they were dosed with Siwutang 10 g · kg^-1 · d^-1 for 7 days, the effects above-mentioned were reversed. Conclusion ： The injury established by injecting CTX is a kind of proper model to develop further metabonomics researches. The damage of mitoehondria, disorder of energy metabolism and osmoregulation are observed in cyclophosphamide caused blood deficiency model by NMR-based-metabonumics method, and the Siwutang can improve these effects.

关 键 词：四物汤 血虚证 环磷酰胺 代谢组学 核磁共振 主成分分析 

分 类 号：R285.5[医药卫生—中药学]