“肝郁证”大鼠模型的建立及代谢组学的初步研究  

Establishment of rat model of syndrome of stagnation of liver qi and primary study using metabonomics

作　　者：徐舒[1] 陈合兵[2] 李洪[1] 张琪[2] 蔡红兵[1] 颜贤忠[2] 吕志平[1] 

机构地区：[1]南方医科大学中医药学院,广州510515 [2]国家生物医学分析中心,北京100850

出　　处：《中华中医药杂志》2009年第6期787-791,共5页China Journal of Traditional Chinese Medicine and Pharmacy

摘　　要：目的:建立并验证"肝郁证"大鼠模型,在此基础上运用代谢组学的方法研究"肝郁证"代谢组学特征及生物标志物,探讨代谢组学用于证候研究的可行性。方法:12只Wistar大鼠随机分为模型组与对照组(每组6只)。模型组采用慢性束缚的方法造成"肝郁证"大鼠模型,通过行为学观察、糖水偏好实验、血浆皮质酮、胃黏膜、肾上腺病理观察评价模型。运用1HNMR技术对两组大鼠血清进行代谢组学分析。结果:模型组大鼠行为异常,主要表现为躲避、活动减少、大便松散、皮色失去光泽,造模第2周开始生长较对照组缓慢(P<0.05);糖水偏好实验显示造模过程中模型组对糖水的偏好摄入少于对照组(P<0.05);造模结束后模型组大鼠血浆皮质酮水平高于对照组(P<0.01);造模后模型组胃黏膜、肾上腺未见明显病理改变,但模型组肾上腺体重指数高于对照组(P<0.05)。代谢组学分析表明,与对照组相比,模型组血浆中葡萄糖(Glucose)和肌酸(Creatine)的含量明显偏高,而3-羟基丁酸(3-HB)、谷氨酰胺(Gln)、磷脂酰胆碱(PtdCho)、磷酸胆碱(PCho)和不饱和脂肪酸(UFA)的含量明显偏低。结论:从病因学、行为学和病理表现上慢性束缚的方法成功构建了肝郁证大鼠模型。代谢组学得出肝郁证模型与对照组大鼠血清代谢组分存在明显差异,这些差异小分子物质构成"肝郁证"的代谢标志物群,成为肝郁证代谢水平的物质基础,有助于阐释肝郁证的本质。代谢组学用于证候研究具有可行性和实用性。Objective： To establish and validate the rat model of syndrome of stagnation of liver qi, followed by a primary study on this model with 1H NMR based on metabonomics to explore the essence of syndrome of stagnation of liver qi. Methods： Twelve Wistar male rats were randomly divided into two groups （model group and control group）. The rats of model group were restrained by special equipment for 21 days to get into stagnation of liver qi. The behavior, fluid consumption test and plasma CORT of rats were recorded. At 22th day, animals were sacrificed and biopsies of gastric mucosa and adrenal gland were collected for pathological check, and serum samples for 1H NMR spectroscopy. The NMR data were analyzed using principal component analysis. Results： There were abnormal behaviors, such as decrease of elusion, slackness, looser stools, and matte fur were observed among model group rats. After one week the body weight of model group was significantly lower than that of control group （P〈0.05）. Fluid consumption test revealed that model rats lost appetite for sweet water （P〈0.05） during the whole procession. The plasma CORT was much higher in model rats than that in normal ones （P〈0.01）. However, no pathological changes were observed in gastric mucosa and adrenal gland of model group rats. Metabonomlc analysis indicated that, compared with control group, blood concentrations of glucose, and creatine increased clearly in the model rats, while 3-HB, Gin, PtdCho, PCho and UFA decreased considerably. Conclusions： Based on etiological, behavioral and pathological observation, it can be concluded that the rat model of syndrome of stagnation of liver qi was successfully established by chronic restrain. Metabonomic analysis can clearly discriminate the metabolic differences between model and normal rats. These differential metabolites can be considered as potential metabolic biomarkers of syndrome of stagnation of liver qi and help explain the essence of it. Metabonomic study based on 1H NMR method

关 键 词：肝郁证 代谢组学 核磁共振 大鼠模型 中医证候 

分 类 号：R-332[医药卫生]