基于代谢组学的逍遥散方证相关的机理研究
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摘要
选题依据:方证对应研究是中医药学研究的核心问题、也是热点和难点问题。目前多以经典方剂为切入点,应用现代技术和相关的分析方法进行尽可能的全方位评价。代谢组学研究机体内源性代谢产物谱的动态变化过程,可以反映出刺激或药物作用的发生、发展和结果的全过程。将其与基因组学和蛋白质组学的结果结合起来,就能够全面、客观地反映刺激或药物作用的情况,为疾病发生发展机理和药理作用机理的研究提供全方位的信息。基于代谢组学强调把人体作为一个完整的系统来研究,通过测定人体各种体液内代谢物的组成变化来认识和反映人体代谢网络在疾病和药物作用下的变化规律,对于揭示复杂性疾病的机理和药物的代谢模式具有独特的优势,也与中医学的整体观、系统观和辨证论治思维非常吻合,与中医重视从人与自然、人与社会和人体内在的普遍联系和动态变化去分析、认识把握疾病发生、发展、变化的客观规律的认识一致。笔者试图以慢性束缚应激大鼠模型(肝郁脾虚证)和逍遥散为切入点,运用代谢组学的技术与方法,探索出一种研究中医方证相应理论的新方法、新途径。
目的:本研究基于代谢组学的研究方法,重点研究基于代谢组学的慢性束缚应激大鼠模型(肝郁脾虚证)的相关变化及逍遥散对其的作用。以NMR,生物信息学等现代技术,通过对血液代谢组的分析,研究慢性束缚和逍遥散干预引起的大鼠内源性代谢物的变化,获得慢性束缚应激(肝郁脾虚证)和逍遥散给药前后的代谢物指纹图谱;分析血液代谢指纹图谱变化的原因,确定与慢性束缚应激(肝郁脾虚证)发生和逍遥散作用机理密切相关的代谢组学特征和小分子标志化合物;阐明药物作用靶点及作用机理,为慢性束缚应激(肝郁脾虚证)和逍遥散作用机理的系统研究提供科学依据。也试图以逍遥散为切入点,运用代谢组学的技术与方法,探索出一种研究中医方证相应理论的新方法、新途径。
方法:选用实验用二级雄性SD大鼠42只,质量200 20g,以慢性束缚方法制作应激大鼠模型,随机分为:7天正常对照组(A组,n=6)、21天正常对照组(B组,n=6)、7天模型组(C组,n=6)、21天模型组(D组,n=6)、7天逍遥散1号组(E组,n=6)、21天逍遥散1号组(F组,n=6)、21天逍遥散2号组(G组,n=6)7组。分别于第8天、第22天,麻醉心室取血。在VARIAN UNITYINOVA600MHz超导傅立叶变化核磁共振波谱仪上分别调用弛豫编辑脉冲序列(CPMG)、扩散编辑脉冲序列(LED)进行实验,采用预饱和方式抑制水峰,饱和时间为2s,谱宽8000Hz,采样点数32k,累加次数64次,预饱和频率和中心频率都在水峰位置。自由感应衰减(FID)信号经过32k点傅立叶变换得到一维NMR谱图。以TSP为化学位移参考峰的位置,设为0ppm。调用VNMR软件中的程序将1H谱中从4.5-0.5ppm (CPMG)以及6.0-0ppm(LED)范围内的谱峰,按每段为0.04ppm,进行分段积分。将积分数据归一化之后,以文本文件或Excel文件贮存,用于模式识别分析。将积分值进行中心化和定标(centeringandscaling),用SIMCA-P 10.0软件包(瑞典,Umetrics AB,Umea)进行主成分分析(PCA),必要时进行判别分析(PLS-DA)。
结果:①A、B、C、D、E、F、G各组间进行的PCA分析结果显示,各样本基本集中分布于得分图的椭圆形(95%置信区内)的4个区域,虽然A、B、C、D、E、F、G各组动物组内样品存在一定的差异,表现为有些样品相对离散,但各对照组间基本仍能分开,说明不同组别之间存在代谢产物的差异。正常组(A组、B组)与模型组(C组、D组)之间代谢产物有明显的不同,说明动物模型造模基本成功,模型组与正常组比较,两者存在代谢网路的改变。模型组(C组、D组)与治疗组(E组、F组、G组)各对照组间基本能分开,说明不同组别之间代谢产物存在差异、代谢网路有所不同,给与逍遥散后可以干预代谢物或代谢途径而致代谢终产物的改变。②正常组与模型组组间代谢标志物比较发现:A、C两组:A组含量高于C组有乳酸(1.36、1.38、1.4、4.16、4.12)、丙氨酸(1.44)、脂类化合物(0.92、1.36)、血糖(3-4)、以及谷氨酸(2.4)、VLDL和LDL(0.87、0.91、0.95、0.99)、HDL(0.83)、胆碱(3.23、3.27)、NAC-1(2.07);C组含量高于A组的有NAC-1(2.08)、小分子化合物(1.12)、3.44ppm未知化合物、长链脂肪酸(1.31、1.23、1.27、1.36、1.39)、NAC-2(2.15)以及1.59ppm处未知化合物。B、D两组:B组含量高于D组的有LDL(0.87、0.91、1.27)、不饱和脂肪酸(4-5)、饱和脂肪酸(4-5);D组含量高于B组的有NAC-1(2.07)、NAC-2(2.16)、饱和脂肪酸(1-3)。③模型组与治疗组组间代谢标志物比较发现:C、E组:C组样本中磷脂酰胆碱(3.28)、脂类化合物(0.92、0.96、1.32)、NAC-2(2.15)、NAC-1(2.08、2.11)、小分子氨基酸(1)、VLDL(0.95、1.35、1.39、1.43)、甘油三酯(3.55、3.59)以及1.63ppm3.2和3.16ppm、3.44ppm等未知化合物含量较高;而E组中乳酸(1.4、4.16、4.2)、谷氨酸(2.44)、丙氨酸(1.52)、缬氨酸(1.08)、HDL(0.87、1.21)、不饱和脂肪酸(5.31、5.35)及小分子化合物(1-2)含量较高。与同时点的A组比较,其乳酸(1.4、4.16、4.2)、谷氨酸(2.44)、丙氨酸(1.52)、HDL(0.87、1.21)有所恢复,并有不饱和脂肪酸(5.31、5.35)及小分子化合物(1-2)含量的提升。D、F组:D组样本中乙酸(1.96)、乳酸(4.2、1.4、4.16)、缬氨酸(1.04)、NAC-1(2.08)、NAC-2(2.15)以及小分子化合物(1-2)、化合物(3-4)含量较高;F组样本中苏氨酸(0.92、0.96、1.32、1.36)、丙氨酸(1.48)、谷氨酸(2.44)以及3.44ppm、VLDL和LDL(0.87、0.91、1.27、1.31、1.35、)、胆碱(3.27)以及不饱和脂肪酸(5.31、5.35)等化合物含量较高。与同时点的B组比较,其VLDL和LDL(0.87、0.91、1.27、1.31、1.35、)及不饱和脂肪酸(5.31、5.35)等化合物有所恢复,并有苏氨酸(0.92、0.96、1.32、1.36)、丙氨酸(1.48)、谷氨酸(2.44)、胆碱(3.27)等化合物含量的增加。并随给药时间的延长,其F组样本中3.4ppm、3.8ppm、2.44ppm未知化合物、磷酯酰胆碱(3.28)、胆碱(3.27)以及LDL或HDL(0.87、0.91)含量更高。D、G组:D组样本中血糖(3-4)、乙酸(1.96)、胆碱(3.24)、NAC-2(2.15)、NAC-1(2.08)、3.2ppm、1.48ppm等处所表示的化合物含量较高、脂类化合物含量相对较低;而G组样本中脂类化合物(0.92、0.96)、乳酸(1.36、1.4、4.16)、苏氨酸(1.32)、甲硫氨酸(2.16)、VLDL和LDL(0.87、0.91、1.27、1.31、1.35、)、磷酯酰胆碱(3.23)以及不饱和脂肪酸(1.99、5.31、5.35)等化合物含量较高。④随给药时间的延长, 3.4ppm、3.8ppm、2.44ppm未知化合物、磷酯酰胆碱(3.28)、胆碱(3.27)以及HDL(0.87、0.91)含量更高。⑤F组(逍遥散1号组)与G组(逍遥散2号组)组间代谢标志物比较发现: F、G两组样本中成分类似。相对而言,G组中各化合物含量比F组要高,G组样本中VLDL和LDL(0.87、0.91、1.27、1.31、1.35)、胆碱(3.23)、不饱和脂肪酸(1.99、5.31、5.35)、乳酸(1.4、1.36、4.16)、丙氨酸(1.52)、苏氨酸(1.32)、脂类化合物(0.92、0.96)等化合物含量较高;F组样本中血糖(3-4)、谷氨酸(2.44)、NAC-1(2.08、2.12)、2.12ppm化合物含量较高。
结论:①代谢组学方法是研究中医症候和中医方证相关机理的良好平台。②从代谢组学的角度来看,慢性束缚应激大鼠模型与中医肝郁脾虚证存在对应关联性。③慢性束缚应激大鼠(肝郁脾虚证)代谢表型为乳酸、胆碱、NAC、饱和脂肪酸、血糖上升降,不饱和脂肪酸、HDL、3.44ppm等化合物含量降低。其代谢产物标志物可能是乳酸、胆碱、NAC、饱和脂肪酸、血糖、不饱和脂肪酸、HDL、3.44ppm等化合物。④慢性束缚应激(肝郁脾虚证)发生的代谢终产物的改变以脂类物质更明显,其具体发生的代谢物、代谢终产物或代谢途径的改变还需进一步研究。⑤逍遥散的作用显示出多途径、多靶点、双向调节特点。从代谢组学角度看,逍遥散可使代谢终产物乳酸、胆碱、NAC、饱和脂肪酸、血糖下降,不饱和脂肪酸、HDL、3.44ppm等化合物含量升高,表现为明显的代谢终产物的调节效应。其作用的靶点包含肝、脑、肾、肌肉、胃肠、脂肪组织、红细胞等多脏器组织细胞,其调节主要干扰何种代谢物或代谢途径而改变代谢终产物还需进一步研究确认。而且21天疗程的疗效优于7天疗程。⑥逍遥散中挥发油成分对慢性束缚应激大鼠(肝郁脾虚证)的代谢终产物的调节可能有负面影响。⑦代谢组学方法还可能用于有效药物组份的鉴别和度量。
创新点:①首次用代谢组学方法研究慢性束缚应激(肝郁脾虚证)及逍遥散的作用机理,是方法学的创新。②首次从代谢组学的角度证明慢性束缚应激大鼠模型与中医肝郁脾虚证的对应关联性。进一步发现慢性束缚应激大鼠(肝郁脾虚证)代谢表型为乳酸、胆碱、NAC、饱和脂肪酸、血糖上升降,不饱和脂肪酸、HDL、3.44ppm等化合物含量降低。其代谢产物标志物可能是乳酸、胆碱、NAC、饱和脂肪酸、血糖、不饱和脂肪酸、HDL、3.44ppm等化合物。慢性束缚应激(肝郁脾虚证)发生的代谢终产物的改变以脂类物质更明显。③首次从代谢组学的角度证明逍遥散的作用显示出多途径、多靶点、双向调节特点。逍遥散可使代谢终产物乳酸、胆碱、NAC、饱和脂肪酸、血糖下降,不饱和脂肪酸、HDL、3.44ppm等化合物含量升高,表现为明显的调节代谢终产物的效应。而且21天疗程的疗效优于7天疗程。因此从代谢组学的角度来研究慢性束缚应激(肝郁脾虚证)及逍遥散的作用机理,丰富了慢性束缚应激大鼠模型、中医肝郁脾虚证、中医肝主疏泄的生物学内涵,初步揭示了逍遥散的作用机理。④首次发现逍遥散中挥发油成分对慢性束缚应激大鼠(肝郁脾虚证)的代谢终产物的调节可能有负面影响。证明代谢组学方法还可能用于有效药物组份的鉴别和度量。
RATIONALES: Studies of prescriptions corresponding to syndromes are the core ofresearches in Traditional Chinese Medicine (TCM). These are also hot but difficult researchpoints in this area. Based on traditional prescriptions, thorough evaluations of these studieshave been attempted by the use of modern technology and relevant analyses. Themetabolomic analysis can be used to determine active alterations of endogenous metabolicspectra and subsequently reveal changes in the whole process, including occurrence,development, and consequences in response to stimuli or drug treatment. In addition, incombination with genomic and protein methods, the metabolomic analysis can be used toexamine actions of stimuli or drugs completely and objectively, which provides entireinformation about processes of diseases and mechanisms of pharmacological actions. Inmetabonomics, the human body is studied as an integrated system, in which changes inmetabolic components of various body fluids are examined and used to reflect alterations ofmetabolic networks in the body under conditions of diseases and treatments with drugs. Thus,the metabolomic analysis has unique advantages in studying mechanisms of complexdiseases and metabolic models of drugs. This is consistent with the views of integration,systems, and dialectical diagnosis and therapeutics in TCM. It is also in agreement with theopinions of understanding and analyzing objective profiles of diseases in their occurrence,development, and alterations in TCM, which emphasizes on the common links of human tonature and societies and connections within the human body. Using metabolomic techniquesand methods, the author found a novel method for studying prescriptions and syndromes inTCM based on the therapeutic effects of the Xiaoyaosan decoction (XYS) on changesinduced by chronic immobilization stress (CIS), which is a model of Liver Stagnation andSpleen Deficiency Syndromes (LSSDS) in rats.
OBJECTIVES: To determine CIS (LSSDS)-induced changes in metabolisms with andwithout XYS treatment using metabolomic analyses. Specifically, we aimed to studyCIS-induced changes in endogenous metabolites and obtain“fingerprint spectra ofmetabolites”under CIS (LSSDS) in the absence and presence of XYS by analyzingmetabolisms in the rat blood using modern techniques such as NMR and bioinformatics; todetermine small molecule marking compounds and“characteristics of metabonomics”,which were related to the mechanisms of CIS (LSSDS) and XYS actions, via analysis ofcauses of changes in blood“fingerprint spectra of metabolites”; to explore drug targets andelucidate mechanisms underlying drug actions, which were expected to provide scientificevidence for systemic studies of CIS (LSSDS) and mechanisms of XYS actions. In addition,novel methods and pathways were anticipated for studies of TCM prescriptions andsyndromes, based on the exploration of XYS using metabolomic techniques and methods.
METHODS: Forty-two male Sprague-Dawley rats, weighing 200±20 g, were subjected to chronic immobilization to develop stress models. Rats were randomly dividedinto 7 groups with 6 rats each: (A) 7-day control, (B) 21-day control, (C) 7-day model(CIS), (D) 21-day model, (E) 7-day XYS-1, (F) 21-day XYS-1, (G) 21-day XYS-2. Bloodwas collected from the cardioventricle under anesthesia on the 8th (A, C and E) or 22nd day(B, D, F and G) and detected CPMG and LED using the Fourier variable superconductingnuclear magnetic resonance (NMR) spectrometer (VARIAN UNITYINOVA 600MHz).Saturated inhibition of the water peak was used, with the saturation time 2 s, spectral width8,000 Hz, sample collecting points 32 k, and accumulation times 64. Both pre-saturated andcentral frequencies were located in the water peak. Free induction decay (FID) signals weretransferred into one-dimensional NMR spectrogram via 32 k Fourier transformation. Thechemical migration reference peak was set to 0 ppm based on TSP. Segmental integralcalculus (0.04 ppm per segment) was performed from 4.5-0.5 ppm (CPMG) and 6.0-0 ppm(LED) within the peak ranges in 1H spectra using the VNMR software. Data were saved astext or Excel files after normalization and then used for pattern recognition analyses.Values by calculus were centering and scaling before PCA or PLS-DA, if necessary, usingthe SIMCA-P10.0 software (Umetrics AB, Umea, Sweden).
RESULTS: (1) Based on the results from PCA in the groups of A-G, the samples wereprimarily distributed in four regions of the scoring oval (95% confidence intervals). Whilevariability existed within each of the groups, as evidenced by relative scatters in somesamples, control groups were basically separated, suggesting differences of metabolitesbetween groups. The metabolites of the model groups (C and D) were significantly differentfrom those of controls (A and B), suggesting that the animal models were successfullyestablished and that they displayed changes in the network of metabolisms relative tocontrols. Models (C and D) and XYS treatment (E, F, and G) were separated from therespective controls, suggesting differences of metabolites and metabolic networks betweengroups; treatment with XYS results in changes in final metabolites via interruptingmetabolites or the pathway of metabolisms. (2) By comparison of metabolic markersbetween controls and models, it was found that, relative to the 7-day CIS model rats (C),non-stress rats (A) displayed higher concentrations of lactic acid (1.36, 1.38, 1.4, 4.16, 4.12),alanine(1.44), lipids (0.92, 1.36), blood sugar(3-4), glutamate(2.4), VLDL, LDL(0.87, 0.91, 0.95, 0.99), HDL(0.83), choline(3.23, 3.27), and NAC-1(2.07)and lowerconcentrations of NAC-1(2.08), small molecule compounds(1.12), long fatty acid(1.31,1.23, 1.27, 1.36, 1.39), NAC-2(2.15), and 1.59 and 3.44 ppm unknown compounds.Similarly, compared to 21-day model rats (D), 21-day stress rats (B) displayed higher levelsof LDL(0.87, 0.91, 1.27), unsaturated fatty acid(4-5), saturated fatty acid (4-5), but lowerlevels of NAC-1 (2.07), NAC-2 (2.16), saturated fatty acid (1-3). (3) By comparison ofmetabolic markers between models and XYS treatment, it was found that CIS rats withoutXYS (C) displayed higher levels of phosphatidylcholine(3.28), lipids(0.92, 0.96, 1.32),NAC-2(2.15), NAC-1(2.08, 2.11), small molecular amino acid (1), VLDL(0.95, 1.35,1.39, 1.43), triglyceride(3.55、3.59), and several unknown compounds (1.63, 3.2, 3.16, and3.44 ppm). By contrast, XYS rats (E) showed higher levels of lactic acid(1.4, 4.16, 4.2), glutamate(2.44), alanine(1.52), valine(1.08), HDL(0.87, 1.21), unsaturated fatty acid(5.31, 5.35), and small molecular compounds (1-2). Compared to the non-stress control(A), XYS resumed lactic acid(1.4, 4.16, 4.2), glutamate(2.44), alanine(1.52), and HDL(0.87, 1.21)to a certain extent; XYS also increased, unsaturated fatty acid(5.31, 5.35), andsmall molecular compounds (1-2). In addition, 21-day model rats (D) exhibited higher levelsof acetic acid (1.9), lactic acid (4.2, 1.4, 4.16), valine(1.04), NAC-1(2.08), NAC-2(2.15)small molecular compounds (1-2), and compounds (3-4). In the presence of XYS (F), the21-day model rats displayed higher levels of threonine(0.92, 0.96, 1.32, 1.36), alanine(1.48), glutamate(2.44), 3.44ppm, VLDL and LDL(0.87, 0.91, 1.27, 1.31, 1.35), choline(3.27), and unsaturated fatty acid(5.31, 5.35). Compared to the 21-day control, XYS-1 inthe F group resumed VLDL and LDL(0.87, 0.91, 1.27, 1.31, 1.35)as well as unsaturatedfatty acid(5.31, 5.35); it also increased threonine(0.92, 0.96, 1.32, 1.36), alanine(1.48),glutamat(e2.44), cholin(e3.27). The levels of unknown compounds (3.4, 3.8, and 2.44ppm),phosphatidylcholine(3.28), choline(3.27), and LDL or HDL(0.87, 0.91)were furtherincreased in the presence of XYS over time. While the 21-day model rats (D) displayedhigher concentrations of blood sugar(3-4), acetic acid(1.96), choline(3.24), NAC-2(2.15)、NAC-1(2.08), and unknown compounds 3.2ppm and1.48ppm, the concentraionof lipids was relatively low. By contrast, the presence of XYS-2 (G) increased lipids(0.92,0.96), lactic acid(1.36, 1.4, 4.16), threonine(1.32), methionine(2.16), VLDL and LDL(0.87, 0.91, 1.27, 1.31, 1.35、), phosphatidylcholin(e3.23), and unsaturated fatty acid(1.99,5.31, 5.35). (4) The contents of the unknown compounds (3.4, 3.8, and 2.44 ppm),phosphatidylcholine(3.28), choline(3.27), and HDL(0.87, 0.91)were further increasedin the presence of XYS over time. (5) Comparison of the metabolic markers between XYS-1(F) and XYS-2 (G) revealed that both treatments affect the components similarly, althoughthe latter increased the contents of compounds to a greater extent relative to the former.Specifically, the XYS-2 rats (G) displayed higher contents of VLDL and LDL(0.87, 0.91,1.27, 1.31, 1.35), choline(3.23), unsaturated fatty acid(1.99, 5.31, 5.35), lactic acid(1.4,1.36, 4.16), alanine(1.52), threonine(1.32), and lipids (0.92, 0.96), whereas the XYS-1rats (F) displayed higher levels of blood suga(r3-4), glutamate(2.44), NAC-1(2.08, 2.12),and the unknown compound 2.12ppm.
CONCLUSIONS: (1) Metabonomics is a useful platform for studying mechanisms ofsymptoms, syndromes, diagnoses, and prescriptions or treatments in TCM. (2) Inmetabolomics, the CIS model has a corresponsive connection with LSSDS in TCM. (3) Themetabolic phenotype of CIS (LSSDS) includes increases in the levels of lactic acid, choline,NAC, saturated fatty acid, and blood sugar and decreases in the levels of unsaturated fattyacid, HDL, and the unknown compound 3.55 ppm. These metabolic components also may bethe markers of metabolites. (4) The final metabolites changed by CIS (LSSDS) primarilyinclude lipids. Further studies are still needed to determine specific metabolites, final metabolites, and changes in metabolic pathways. (5) The roles of XYS are characteristic ofmulti-pathways, multi-targets, and bi-direction regulation. In the view of metabonomics,XYS markedly regulates the contents of final metabolites, as evidenced by decreased lacticacid, choline, NAC, saturated fatty acid, and blood sugar and increased unsaturated fatty acid,HDL, and the compound 3.44 ppm. The targets of XYS include various organs, tissues, andcells, such as the liver, brain, kidney, muscles, gastrointestine, fatty tissues, and red bloodcells. Further studies are needed to determine which metabolites or metabolic pathways XYSinterrupts to alter final metabolites. The efficacy of 21-day XYS is greater than that of 7-dayXYS. (6) The vapor oil in XYS may play a negative role in the regulation of finalmetabolites by CIS (LSSDS). (7) Metabonomics can also be used to distinct and quantifyeffective components of medicines.
INNOVATIONS: (1) Metabolomic analyses were used for the first time to study themechanisms underlying CIS (LSSDS) and XYS actions; this was an innovation inmethodology. (2) It was demonstrated metabolomically for the first time that CIS wascorrespondingly associated with LSSDS. It was found for the first time that the metabolicphenotype of CIS (LSSDS) includes increases in lactic acid, choline, NAC, saturated fattyacid, and blood sugar and decreases in unsaturated fatty acid, HDL, and the unknowncompound 3.55 ppm. These metabolic components also may be the markers of metabolites.CIS (LSSDS)-induced alterations of final metabolites primarily include lipids. (3) It wasdemonstrated metabolomically for the first time that the roles of XYS are characteristic ofmulti-pathways, multi-targets, and bi-direction regulation. XYS markedly regulates thecontents of final metabolites, as evidenced by decreased lactic acid, choline, NAC, saturatedfatty acid, and blood sugar and increased unsaturated fatty acid, HDL, and the compound3.44 ppm. The potency of 21-day XYS treatment was greater than that of 7-day XYS.Mechanisms underlying CIS (LSSDS) and XYS actions were examined metabolomically forthe first time. This enriched biological contents of CIS rat models, TCM LSSDS, andliver-dominated dispersion and discharge and revealed initial mechanisms of XYS actions.(4) It was found for the first time that the vapor oil in XYS might play a negative role in theregulation of final metabolites by CIS (LSSDS). It demonstrated that metabolomicalmethods could also be used to distinct and quantify effective components of medicines.
目的:本研究基于代谢组学的研究方法,重点研究基于代谢组学的慢性束缚应激大鼠模型(肝郁脾虚证)的相关变化及逍遥散对其的作用。以NMR,生物信息学等现代技术,通过对血液代谢组的分析,研究慢性束缚和逍遥散干预引起的大鼠内源性代谢物的变化,获得慢性束缚应激(肝郁脾虚证)和逍遥散给药前后的代谢物指纹图谱;分析血液代谢指纹图谱变化的原因,确定与慢性束缚应激(肝郁脾虚证)发生和逍遥散作用机理密切相关的代谢组学特征和小分子标志化合物;阐明药物作用靶点及作用机理,为慢性束缚应激(肝郁脾虚证)和逍遥散作用机理的系统研究提供科学依据。也试图以逍遥散为切入点,运用代谢组学的技术与方法,探索出一种研究中医方证相应理论的新方法、新途径。
方法:选用实验用二级雄性SD大鼠42只,质量200 20g,以慢性束缚方法制作应激大鼠模型,随机分为:7天正常对照组(A组,n=6)、21天正常对照组(B组,n=6)、7天模型组(C组,n=6)、21天模型组(D组,n=6)、7天逍遥散1号组(E组,n=6)、21天逍遥散1号组(F组,n=6)、21天逍遥散2号组(G组,n=6)7组。分别于第8天、第22天,麻醉心室取血。在VARIAN UNITYINOVA600MHz超导傅立叶变化核磁共振波谱仪上分别调用弛豫编辑脉冲序列(CPMG)、扩散编辑脉冲序列(LED)进行实验,采用预饱和方式抑制水峰,饱和时间为2s,谱宽8000Hz,采样点数32k,累加次数64次,预饱和频率和中心频率都在水峰位置。自由感应衰减(FID)信号经过32k点傅立叶变换得到一维NMR谱图。以TSP为化学位移参考峰的位置,设为0ppm。调用VNMR软件中的程序将1H谱中从4.5-0.5ppm (CPMG)以及6.0-0ppm(LED)范围内的谱峰,按每段为0.04ppm,进行分段积分。将积分数据归一化之后,以文本文件或Excel文件贮存,用于模式识别分析。将积分值进行中心化和定标(centeringandscaling),用SIMCA-P 10.0软件包(瑞典,Umetrics AB,Umea)进行主成分分析(PCA),必要时进行判别分析(PLS-DA)。
结果:①A、B、C、D、E、F、G各组间进行的PCA分析结果显示,各样本基本集中分布于得分图的椭圆形(95%置信区内)的4个区域,虽然A、B、C、D、E、F、G各组动物组内样品存在一定的差异,表现为有些样品相对离散,但各对照组间基本仍能分开,说明不同组别之间存在代谢产物的差异。正常组(A组、B组)与模型组(C组、D组)之间代谢产物有明显的不同,说明动物模型造模基本成功,模型组与正常组比较,两者存在代谢网路的改变。模型组(C组、D组)与治疗组(E组、F组、G组)各对照组间基本能分开,说明不同组别之间代谢产物存在差异、代谢网路有所不同,给与逍遥散后可以干预代谢物或代谢途径而致代谢终产物的改变。②正常组与模型组组间代谢标志物比较发现:A、C两组:A组含量高于C组有乳酸(1.36、1.38、1.4、4.16、4.12)、丙氨酸(1.44)、脂类化合物(0.92、1.36)、血糖(3-4)、以及谷氨酸(2.4)、VLDL和LDL(0.87、0.91、0.95、0.99)、HDL(0.83)、胆碱(3.23、3.27)、NAC-1(2.07);C组含量高于A组的有NAC-1(2.08)、小分子化合物(1.12)、3.44ppm未知化合物、长链脂肪酸(1.31、1.23、1.27、1.36、1.39)、NAC-2(2.15)以及1.59ppm处未知化合物。B、D两组:B组含量高于D组的有LDL(0.87、0.91、1.27)、不饱和脂肪酸(4-5)、饱和脂肪酸(4-5);D组含量高于B组的有NAC-1(2.07)、NAC-2(2.16)、饱和脂肪酸(1-3)。③模型组与治疗组组间代谢标志物比较发现:C、E组:C组样本中磷脂酰胆碱(3.28)、脂类化合物(0.92、0.96、1.32)、NAC-2(2.15)、NAC-1(2.08、2.11)、小分子氨基酸(1)、VLDL(0.95、1.35、1.39、1.43)、甘油三酯(3.55、3.59)以及1.63ppm3.2和3.16ppm、3.44ppm等未知化合物含量较高;而E组中乳酸(1.4、4.16、4.2)、谷氨酸(2.44)、丙氨酸(1.52)、缬氨酸(1.08)、HDL(0.87、1.21)、不饱和脂肪酸(5.31、5.35)及小分子化合物(1-2)含量较高。与同时点的A组比较,其乳酸(1.4、4.16、4.2)、谷氨酸(2.44)、丙氨酸(1.52)、HDL(0.87、1.21)有所恢复,并有不饱和脂肪酸(5.31、5.35)及小分子化合物(1-2)含量的提升。D、F组:D组样本中乙酸(1.96)、乳酸(4.2、1.4、4.16)、缬氨酸(1.04)、NAC-1(2.08)、NAC-2(2.15)以及小分子化合物(1-2)、化合物(3-4)含量较高;F组样本中苏氨酸(0.92、0.96、1.32、1.36)、丙氨酸(1.48)、谷氨酸(2.44)以及3.44ppm、VLDL和LDL(0.87、0.91、1.27、1.31、1.35、)、胆碱(3.27)以及不饱和脂肪酸(5.31、5.35)等化合物含量较高。与同时点的B组比较,其VLDL和LDL(0.87、0.91、1.27、1.31、1.35、)及不饱和脂肪酸(5.31、5.35)等化合物有所恢复,并有苏氨酸(0.92、0.96、1.32、1.36)、丙氨酸(1.48)、谷氨酸(2.44)、胆碱(3.27)等化合物含量的增加。并随给药时间的延长,其F组样本中3.4ppm、3.8ppm、2.44ppm未知化合物、磷酯酰胆碱(3.28)、胆碱(3.27)以及LDL或HDL(0.87、0.91)含量更高。D、G组:D组样本中血糖(3-4)、乙酸(1.96)、胆碱(3.24)、NAC-2(2.15)、NAC-1(2.08)、3.2ppm、1.48ppm等处所表示的化合物含量较高、脂类化合物含量相对较低;而G组样本中脂类化合物(0.92、0.96)、乳酸(1.36、1.4、4.16)、苏氨酸(1.32)、甲硫氨酸(2.16)、VLDL和LDL(0.87、0.91、1.27、1.31、1.35、)、磷酯酰胆碱(3.23)以及不饱和脂肪酸(1.99、5.31、5.35)等化合物含量较高。④随给药时间的延长, 3.4ppm、3.8ppm、2.44ppm未知化合物、磷酯酰胆碱(3.28)、胆碱(3.27)以及HDL(0.87、0.91)含量更高。⑤F组(逍遥散1号组)与G组(逍遥散2号组)组间代谢标志物比较发现: F、G两组样本中成分类似。相对而言,G组中各化合物含量比F组要高,G组样本中VLDL和LDL(0.87、0.91、1.27、1.31、1.35)、胆碱(3.23)、不饱和脂肪酸(1.99、5.31、5.35)、乳酸(1.4、1.36、4.16)、丙氨酸(1.52)、苏氨酸(1.32)、脂类化合物(0.92、0.96)等化合物含量较高;F组样本中血糖(3-4)、谷氨酸(2.44)、NAC-1(2.08、2.12)、2.12ppm化合物含量较高。
结论:①代谢组学方法是研究中医症候和中医方证相关机理的良好平台。②从代谢组学的角度来看,慢性束缚应激大鼠模型与中医肝郁脾虚证存在对应关联性。③慢性束缚应激大鼠(肝郁脾虚证)代谢表型为乳酸、胆碱、NAC、饱和脂肪酸、血糖上升降,不饱和脂肪酸、HDL、3.44ppm等化合物含量降低。其代谢产物标志物可能是乳酸、胆碱、NAC、饱和脂肪酸、血糖、不饱和脂肪酸、HDL、3.44ppm等化合物。④慢性束缚应激(肝郁脾虚证)发生的代谢终产物的改变以脂类物质更明显,其具体发生的代谢物、代谢终产物或代谢途径的改变还需进一步研究。⑤逍遥散的作用显示出多途径、多靶点、双向调节特点。从代谢组学角度看,逍遥散可使代谢终产物乳酸、胆碱、NAC、饱和脂肪酸、血糖下降,不饱和脂肪酸、HDL、3.44ppm等化合物含量升高,表现为明显的代谢终产物的调节效应。其作用的靶点包含肝、脑、肾、肌肉、胃肠、脂肪组织、红细胞等多脏器组织细胞,其调节主要干扰何种代谢物或代谢途径而改变代谢终产物还需进一步研究确认。而且21天疗程的疗效优于7天疗程。⑥逍遥散中挥发油成分对慢性束缚应激大鼠(肝郁脾虚证)的代谢终产物的调节可能有负面影响。⑦代谢组学方法还可能用于有效药物组份的鉴别和度量。
创新点:①首次用代谢组学方法研究慢性束缚应激(肝郁脾虚证)及逍遥散的作用机理,是方法学的创新。②首次从代谢组学的角度证明慢性束缚应激大鼠模型与中医肝郁脾虚证的对应关联性。进一步发现慢性束缚应激大鼠(肝郁脾虚证)代谢表型为乳酸、胆碱、NAC、饱和脂肪酸、血糖上升降,不饱和脂肪酸、HDL、3.44ppm等化合物含量降低。其代谢产物标志物可能是乳酸、胆碱、NAC、饱和脂肪酸、血糖、不饱和脂肪酸、HDL、3.44ppm等化合物。慢性束缚应激(肝郁脾虚证)发生的代谢终产物的改变以脂类物质更明显。③首次从代谢组学的角度证明逍遥散的作用显示出多途径、多靶点、双向调节特点。逍遥散可使代谢终产物乳酸、胆碱、NAC、饱和脂肪酸、血糖下降,不饱和脂肪酸、HDL、3.44ppm等化合物含量升高,表现为明显的调节代谢终产物的效应。而且21天疗程的疗效优于7天疗程。因此从代谢组学的角度来研究慢性束缚应激(肝郁脾虚证)及逍遥散的作用机理,丰富了慢性束缚应激大鼠模型、中医肝郁脾虚证、中医肝主疏泄的生物学内涵,初步揭示了逍遥散的作用机理。④首次发现逍遥散中挥发油成分对慢性束缚应激大鼠(肝郁脾虚证)的代谢终产物的调节可能有负面影响。证明代谢组学方法还可能用于有效药物组份的鉴别和度量。
RATIONALES: Studies of prescriptions corresponding to syndromes are the core ofresearches in Traditional Chinese Medicine (TCM). These are also hot but difficult researchpoints in this area. Based on traditional prescriptions, thorough evaluations of these studieshave been attempted by the use of modern technology and relevant analyses. Themetabolomic analysis can be used to determine active alterations of endogenous metabolicspectra and subsequently reveal changes in the whole process, including occurrence,development, and consequences in response to stimuli or drug treatment. In addition, incombination with genomic and protein methods, the metabolomic analysis can be used toexamine actions of stimuli or drugs completely and objectively, which provides entireinformation about processes of diseases and mechanisms of pharmacological actions. Inmetabonomics, the human body is studied as an integrated system, in which changes inmetabolic components of various body fluids are examined and used to reflect alterations ofmetabolic networks in the body under conditions of diseases and treatments with drugs. Thus,the metabolomic analysis has unique advantages in studying mechanisms of complexdiseases and metabolic models of drugs. This is consistent with the views of integration,systems, and dialectical diagnosis and therapeutics in TCM. It is also in agreement with theopinions of understanding and analyzing objective profiles of diseases in their occurrence,development, and alterations in TCM, which emphasizes on the common links of human tonature and societies and connections within the human body. Using metabolomic techniquesand methods, the author found a novel method for studying prescriptions and syndromes inTCM based on the therapeutic effects of the Xiaoyaosan decoction (XYS) on changesinduced by chronic immobilization stress (CIS), which is a model of Liver Stagnation andSpleen Deficiency Syndromes (LSSDS) in rats.
OBJECTIVES: To determine CIS (LSSDS)-induced changes in metabolisms with andwithout XYS treatment using metabolomic analyses. Specifically, we aimed to studyCIS-induced changes in endogenous metabolites and obtain“fingerprint spectra ofmetabolites”under CIS (LSSDS) in the absence and presence of XYS by analyzingmetabolisms in the rat blood using modern techniques such as NMR and bioinformatics; todetermine small molecule marking compounds and“characteristics of metabonomics”,which were related to the mechanisms of CIS (LSSDS) and XYS actions, via analysis ofcauses of changes in blood“fingerprint spectra of metabolites”; to explore drug targets andelucidate mechanisms underlying drug actions, which were expected to provide scientificevidence for systemic studies of CIS (LSSDS) and mechanisms of XYS actions. In addition,novel methods and pathways were anticipated for studies of TCM prescriptions andsyndromes, based on the exploration of XYS using metabolomic techniques and methods.
METHODS: Forty-two male Sprague-Dawley rats, weighing 200±20 g, were subjected to chronic immobilization to develop stress models. Rats were randomly dividedinto 7 groups with 6 rats each: (A) 7-day control, (B) 21-day control, (C) 7-day model(CIS), (D) 21-day model, (E) 7-day XYS-1, (F) 21-day XYS-1, (G) 21-day XYS-2. Bloodwas collected from the cardioventricle under anesthesia on the 8th (A, C and E) or 22nd day(B, D, F and G) and detected CPMG and LED using the Fourier variable superconductingnuclear magnetic resonance (NMR) spectrometer (VARIAN UNITYINOVA 600MHz).Saturated inhibition of the water peak was used, with the saturation time 2 s, spectral width8,000 Hz, sample collecting points 32 k, and accumulation times 64. Both pre-saturated andcentral frequencies were located in the water peak. Free induction decay (FID) signals weretransferred into one-dimensional NMR spectrogram via 32 k Fourier transformation. Thechemical migration reference peak was set to 0 ppm based on TSP. Segmental integralcalculus (0.04 ppm per segment) was performed from 4.5-0.5 ppm (CPMG) and 6.0-0 ppm(LED) within the peak ranges in 1H spectra using the VNMR software. Data were saved astext or Excel files after normalization and then used for pattern recognition analyses.Values by calculus were centering and scaling before PCA or PLS-DA, if necessary, usingthe SIMCA-P10.0 software (Umetrics AB, Umea, Sweden).
RESULTS: (1) Based on the results from PCA in the groups of A-G, the samples wereprimarily distributed in four regions of the scoring oval (95% confidence intervals). Whilevariability existed within each of the groups, as evidenced by relative scatters in somesamples, control groups were basically separated, suggesting differences of metabolitesbetween groups. The metabolites of the model groups (C and D) were significantly differentfrom those of controls (A and B), suggesting that the animal models were successfullyestablished and that they displayed changes in the network of metabolisms relative tocontrols. Models (C and D) and XYS treatment (E, F, and G) were separated from therespective controls, suggesting differences of metabolites and metabolic networks betweengroups; treatment with XYS results in changes in final metabolites via interruptingmetabolites or the pathway of metabolisms. (2) By comparison of metabolic markersbetween controls and models, it was found that, relative to the 7-day CIS model rats (C),non-stress rats (A) displayed higher concentrations of lactic acid (1.36, 1.38, 1.4, 4.16, 4.12),alanine(1.44), lipids (0.92, 1.36), blood sugar(3-4), glutamate(2.4), VLDL, LDL(0.87, 0.91, 0.95, 0.99), HDL(0.83), choline(3.23, 3.27), and NAC-1(2.07)and lowerconcentrations of NAC-1(2.08), small molecule compounds(1.12), long fatty acid(1.31,1.23, 1.27, 1.36, 1.39), NAC-2(2.15), and 1.59 and 3.44 ppm unknown compounds.Similarly, compared to 21-day model rats (D), 21-day stress rats (B) displayed higher levelsof LDL(0.87, 0.91, 1.27), unsaturated fatty acid(4-5), saturated fatty acid (4-5), but lowerlevels of NAC-1 (2.07), NAC-2 (2.16), saturated fatty acid (1-3). (3) By comparison ofmetabolic markers between models and XYS treatment, it was found that CIS rats withoutXYS (C) displayed higher levels of phosphatidylcholine(3.28), lipids(0.92, 0.96, 1.32),NAC-2(2.15), NAC-1(2.08, 2.11), small molecular amino acid (1), VLDL(0.95, 1.35,1.39, 1.43), triglyceride(3.55、3.59), and several unknown compounds (1.63, 3.2, 3.16, and3.44 ppm). By contrast, XYS rats (E) showed higher levels of lactic acid(1.4, 4.16, 4.2), glutamate(2.44), alanine(1.52), valine(1.08), HDL(0.87, 1.21), unsaturated fatty acid(5.31, 5.35), and small molecular compounds (1-2). Compared to the non-stress control(A), XYS resumed lactic acid(1.4, 4.16, 4.2), glutamate(2.44), alanine(1.52), and HDL(0.87, 1.21)to a certain extent; XYS also increased, unsaturated fatty acid(5.31, 5.35), andsmall molecular compounds (1-2). In addition, 21-day model rats (D) exhibited higher levelsof acetic acid (1.9), lactic acid (4.2, 1.4, 4.16), valine(1.04), NAC-1(2.08), NAC-2(2.15)small molecular compounds (1-2), and compounds (3-4). In the presence of XYS (F), the21-day model rats displayed higher levels of threonine(0.92, 0.96, 1.32, 1.36), alanine(1.48), glutamate(2.44), 3.44ppm, VLDL and LDL(0.87, 0.91, 1.27, 1.31, 1.35), choline(3.27), and unsaturated fatty acid(5.31, 5.35). Compared to the 21-day control, XYS-1 inthe F group resumed VLDL and LDL(0.87, 0.91, 1.27, 1.31, 1.35)as well as unsaturatedfatty acid(5.31, 5.35); it also increased threonine(0.92, 0.96, 1.32, 1.36), alanine(1.48),glutamat(e2.44), cholin(e3.27). The levels of unknown compounds (3.4, 3.8, and 2.44ppm),phosphatidylcholine(3.28), choline(3.27), and LDL or HDL(0.87, 0.91)were furtherincreased in the presence of XYS over time. While the 21-day model rats (D) displayedhigher concentrations of blood sugar(3-4), acetic acid(1.96), choline(3.24), NAC-2(2.15)、NAC-1(2.08), and unknown compounds 3.2ppm and1.48ppm, the concentraionof lipids was relatively low. By contrast, the presence of XYS-2 (G) increased lipids(0.92,0.96), lactic acid(1.36, 1.4, 4.16), threonine(1.32), methionine(2.16), VLDL and LDL(0.87, 0.91, 1.27, 1.31, 1.35、), phosphatidylcholin(e3.23), and unsaturated fatty acid(1.99,5.31, 5.35). (4) The contents of the unknown compounds (3.4, 3.8, and 2.44 ppm),phosphatidylcholine(3.28), choline(3.27), and HDL(0.87, 0.91)were further increasedin the presence of XYS over time. (5) Comparison of the metabolic markers between XYS-1(F) and XYS-2 (G) revealed that both treatments affect the components similarly, althoughthe latter increased the contents of compounds to a greater extent relative to the former.Specifically, the XYS-2 rats (G) displayed higher contents of VLDL and LDL(0.87, 0.91,1.27, 1.31, 1.35), choline(3.23), unsaturated fatty acid(1.99, 5.31, 5.35), lactic acid(1.4,1.36, 4.16), alanine(1.52), threonine(1.32), and lipids (0.92, 0.96), whereas the XYS-1rats (F) displayed higher levels of blood suga(r3-4), glutamate(2.44), NAC-1(2.08, 2.12),and the unknown compound 2.12ppm.
CONCLUSIONS: (1) Metabonomics is a useful platform for studying mechanisms ofsymptoms, syndromes, diagnoses, and prescriptions or treatments in TCM. (2) Inmetabolomics, the CIS model has a corresponsive connection with LSSDS in TCM. (3) Themetabolic phenotype of CIS (LSSDS) includes increases in the levels of lactic acid, choline,NAC, saturated fatty acid, and blood sugar and decreases in the levels of unsaturated fattyacid, HDL, and the unknown compound 3.55 ppm. These metabolic components also may bethe markers of metabolites. (4) The final metabolites changed by CIS (LSSDS) primarilyinclude lipids. Further studies are still needed to determine specific metabolites, final metabolites, and changes in metabolic pathways. (5) The roles of XYS are characteristic ofmulti-pathways, multi-targets, and bi-direction regulation. In the view of metabonomics,XYS markedly regulates the contents of final metabolites, as evidenced by decreased lacticacid, choline, NAC, saturated fatty acid, and blood sugar and increased unsaturated fatty acid,HDL, and the compound 3.44 ppm. The targets of XYS include various organs, tissues, andcells, such as the liver, brain, kidney, muscles, gastrointestine, fatty tissues, and red bloodcells. Further studies are needed to determine which metabolites or metabolic pathways XYSinterrupts to alter final metabolites. The efficacy of 21-day XYS is greater than that of 7-dayXYS. (6) The vapor oil in XYS may play a negative role in the regulation of finalmetabolites by CIS (LSSDS). (7) Metabonomics can also be used to distinct and quantifyeffective components of medicines.
INNOVATIONS: (1) Metabolomic analyses were used for the first time to study themechanisms underlying CIS (LSSDS) and XYS actions; this was an innovation inmethodology. (2) It was demonstrated metabolomically for the first time that CIS wascorrespondingly associated with LSSDS. It was found for the first time that the metabolicphenotype of CIS (LSSDS) includes increases in lactic acid, choline, NAC, saturated fattyacid, and blood sugar and decreases in unsaturated fatty acid, HDL, and the unknowncompound 3.55 ppm. These metabolic components also may be the markers of metabolites.CIS (LSSDS)-induced alterations of final metabolites primarily include lipids. (3) It wasdemonstrated metabolomically for the first time that the roles of XYS are characteristic ofmulti-pathways, multi-targets, and bi-direction regulation. XYS markedly regulates thecontents of final metabolites, as evidenced by decreased lactic acid, choline, NAC, saturatedfatty acid, and blood sugar and increased unsaturated fatty acid, HDL, and the compound3.44 ppm. The potency of 21-day XYS treatment was greater than that of 7-day XYS.Mechanisms underlying CIS (LSSDS) and XYS actions were examined metabolomically forthe first time. This enriched biological contents of CIS rat models, TCM LSSDS, andliver-dominated dispersion and discharge and revealed initial mechanisms of XYS actions.(4) It was found for the first time that the vapor oil in XYS might play a negative role in theregulation of final metabolites by CIS (LSSDS). It demonstrated that metabolomicalmethods could also be used to distinct and quantify effective components of medicines.
引文
1.黄煌.论方证相关及其意义.中国中医基础医学杂志.1998,4(6):11
2.王阶、张兰凤、王永炎.方证对应理论源流及临床研究.世界科学技术-中医药现代化.2004,6(4):13-17
3.李致重.日本汉方医学衰落轨迹.崔月犁.中医沉思录.北京:中医古籍出版社,1997,162-170.
4.刘渡舟.方证相对论.北京中医药大学学报.1996,19(1):3
5.谢鸣. 方证相关 逻辑命题及其意义.北京中医药大学学报,2003,26(2):11-12
6.李瑞,尹英杰,林殷,鲁兆麟.中医方证客观化研究初探.北京中医药大学学报 2002;25(3):6-10
7.窦志芳,郭 蕾,张俊龙等. 方证对应 研究的思考.中医杂志.2006,47(8):563
8.王阶.方证对应与方证标准规范探讨.中医杂志 ,2002;43(7):489
9.俞雪如.柯琴 以方名证 思想对吉益东洞 方证相对说 的影响.上海中医药大学学报,2001,15(2):2
10.李国臣,王冠民,崔文艺.胡希恕方证辨证说略.上海中医药杂志,2003,37(10):39.
11.邓铁涛.再论辨证论治.新中医,1999,31(4):8
12.金海浩.中医学的方证相应论.黑龙江中医药,2005,1(1):45
13.张兰凤,王 阶,王永炎.方证对应研究.中华中医药杂志,2005,20(1):8
14.刘晋平,黄宇虹,陆小左.数据挖掘在中医脉诊中的应用.天津中医学院学报.2003,22(3):9
15.姚美村,袁月梅,艾路.数据挖掘及其在中医药现代化中的应用.北京中医药大学学报,2002,25(5):20
16.刘娟,蒋永光,胡波等.白术类方的药证关联分析.成都中医药大学学报.2004,27(4):5
17.孙占全,刘艳骄,西广成等.证候与方剂相关性的研究.中国中医基础医学杂志.2006,12(2):150
18.王米渠,张卫,李珉,等.基因芯片筛选针刺衰老鼠涌泉穴的 318个差异表达基因初报 .中国中西医结合杂志,2002,22(11):928.
19.王米渠,冯韧,严石林等.基因表达谱芯片发现热药疗寒的相关七类基因初报.中医杂志,2003,44(4):228
20.高月,刘永学,马增春等.白芍在四物汤中的反佐作用.中国实验方剂学杂志,2004,10(2):26-28
21.郭平,马增春,李鹰飞等.四物汤对放射线致血虚证小鼠骨髓蛋白质表达的影响.中国中药杂志,2004,29(9):893-896
22.梁乾德,路晓钦,马增春等.四物汤促进造血功能成分的初步研究.中国中药杂志,2004,6:546-549
23.马增春,高月,谭洪玲等.四物汤对辐射致血虚证小鼠血清蛋白质表达的影响.中国中药杂志,2003,28(11):1050-1054
24.高月,马增春,刘永学等.四物汤及其提取物对辐射致血虚证小鼠造血作用的研究.天津中医药,2003,6:47-51
25.高月,马增春,刘永学等.CD34+细胞在血虚证小鼠中的变化及意义.中华中西医杂志,2003,4(5):658-659
26.佟丽,陈苏红,马增春等.用寡核苷酸芯片研究血虚小鼠细胞因子基因表达谱.中草药,2003,7:625-629
27.高月,马增春,陈鹏等.四物汤及其各单药对辐射小鼠骨髓造血的调节作用.中药新药与临床药理,2002,12(5):336-339
28.马增春,高月,谭洪玲等.实验性血虚证小鼠骨髓损伤机制的研究.中医杂志,2002,43(增刊):73-74
29.谭洪玲,马增春,高月等.四物汤对血虚证小鼠骨髓细胞CD34抗原表达的影响.中药新药与临床药理,2002,13(1):11-13
30.马淑华,王升启,范保星等.抑制消减杂交和辐射小鼠血虚模型筛选 四物汤 诱导表达的基因.生物技术通讯,2002,13(2):214-217
31.马增春,高月,刘永学等.辐射小鼠骨髓中CD34+细胞的变化及其意义.中华放射医学与防护杂志,2001,21(1):12-15
32.马增春,高月,刘永学等.环磷酰胺所致血虚证小鼠骨髓CD34+细胞的变化及其意义.中国基础医学杂志,2001,7(7):42-44
33.马增春,高月,刘永学等.四物汤对环磷酰胺所致血虚证小鼠造血细胞作用的研究.中国实验方剂学杂志,2001,7(5):13-15
34.马增春,高月,刘永学等.四物汤对 γ 射线照射致血虚证小鼠造血细胞作用的研究.中国实验方剂学杂志,2001,7(3):41-44
35.谭洪玲,高月,马增春等.四物汤刺激 Peyer sPatch促进骨髓造血的研究.中药药理与临床,2001,17(2):1-3
36.路晓钦,马增春,高月.四物汤药理研究进展.中国实验方剂学杂志,2001,7(4):56-69
37.马增春,高月.血虚证的病理研究概况.新中医,2000,32(9):60-61
38.马增春,高月,刘永学等.再生障碍性贫血患者骨髓造血调控机制的研究进展.国外医学输血及血液学分册,2000,5:351-354
39.刘永学,马增春,高月等.实验性血虚证小鼠骨髓造血干/祖细胞的变化.中国基础医学杂志,2000,6(增刊):337-341
40.董波,马增春,高月等.碘化丙啶对小鼠骨髓 CD34细胞计数的影响.中国应用生理学杂志,2000,16(3):133
41.廖建群,钱锋,马增春等.中医补血方剂对受照小鼠造血功能的影响.生物技术通讯,2000,11(3):214
42.Gennan JB,Bauman DE,Burrrin DG,et al.Metabolomics in the opening decade of the21st century:building the roads to individualized health.J Nutn,2004,134(10):2729
43.GavaghanCL,Holmes E,I_enz E,et al .AnNMR-based metabonomic approach to investigate the biochemi calconsequences of genetic strain diferencesi application to the C57BL10J and AIpk:ApfCDmouse[J].FEBSLett,2O00,484(3):169 174
44.赵剑宇,颜贤忠.基于核磁共振的代谢组学研究进展.国外医学药学分册,2004,31(5):308～
31245.沈一丁,费贵强,张 宇.代谢组学促进中医药现代化.精细化工,2005,22(6):444
46.贾 伟,蒋 健,刘 平等.代谢组学在中医药复杂理论体系研究中的应用.中国中药杂志.2006,31(8):621
47.王桐生,谢鸣.代谢组学与中医药现代化.中医杂志.2006,47(10):723-725
48.衷敬柏,王阶,赵宜军等.病证结合与方证相应研究.辽宁中医杂志.2006;33(2):137-139
49.李茹柳,陈艳芬,陈蔚文.左金丸方证相应动物模型建立的回顾及其对方证研究的意义.广州中医药大学学报.2005;22(3):236-238
50.陈艳芬,陈蔚文,李茹柳.左金丸与反左金丸的方证对应实验研究.广东药学院学报2004;20(5):501-503
51.年莉,方玲,刘鸿雁.吴茱萸汤方证学研究.天津中医药.2004;21(1):38-40
52.费忠东.大柴胡汤现代方证研究.江苏中医药.2003;24(8):43-45
53.王阶,荆鲁,衷敬柏.血府逐瘀汤拆方临床研究.中国中药杂志.2004;29(8):803-807
54.张兰凤,王阶,衷敬柏,等.冠心病病证结合方证对应临床研究.中医杂志.2004;45(6):444
55.张杰.利用方证相应学说探寻中医证的物质基础.中国中医药信息杂志.2005;12(11):3-5
56.李沛清,刘喜平.对中医证与方证研究的思考.中国中医药信息杂志.2005;12(4):5-6
57.王建红,罗来成,于润桥.基于系统工程理论的中医方证研究.辽宁中医杂志.2006;33(3):296
58.王文健,沈自尹,张新民,等.补肾法对老年男性下丘脑-垂体-性腺轴作用的临床和实验研究.中医杂志 1986;27(4):32
59.蔡定芳,沈自尹,张玲绢,等.右归饮对大鼠下丘脑-垂体-肾上腺-胸腺轴功能抑制模型的影响.中国免疫学杂志 1994;10(7):236
60.蔡定芳,沈自尹,陈晓红,等.乌头碱对大鼠下丘脑促肾上腺皮质激素释放激素含量的影响.中国中西医结合杂志 1996;16(9):544
61.钟历勇,沈自尹,蔡定芳,等.补肾健脾活血三类复方对下丘脑-垂体-肾上腺-胸腺轴及 CRF基因表达的影响.中国中西医结合杂志 1997;17(1):39
62.俞瑾,陈红英,毛秋芝.肾主生殖的实验研究.中西医结合杂志 1989;9(9):548
63.俞瑾.肾主生殖与生命网络研究中的启示.中国中西医结合杂志 2000;20(6):409
64.路晓钦,高月,梁乾德,谭洪玲.四物汤中不同组分对血虚模型小鼠造血功能的影响.中成药2005;7(10):1189-1193
65.陈家旭,唐已婷.逍遥散对慢性束缚应激模型大鼠相关脑区 CRF基因表达的影响.中国应用生理杂志 2004;20(1):71-74
66.陈家旭,杨建新,赵歆,等.慢性束缚应激大鼠下丘脑-内啡肽变化及中药复方作用的影响.中国医药学报 2004;19(2):42-44
67.嵇波,陈家旭,鲁兆麟,等.逍遥散对人体神经内分泌免疫系统的影响.北京中医药大学学报2003;26(6):72-75
68.马增春,高月,谭洪玲,梁乾德,王升启.用分子中药组学技术研究四物汤补血的作用机理.世界科学技术中医药现代化,2005,7(3):24-28
69.刘昌孝.中药药代动力学研究的难点和热点.药学学报,2005,40(5):395-401李峰,康廷国.系统论与中药复方作用机制探讨.中医药学刊,2003,21(9):1479-1480
70.王本祥,周秋丽.关于中药活性成分的认识及其研究方法.中国中药杂志,2001,26(1):10-13
71.潘卫松,刘美风,石钺,等.血清药理学、血清化学和中药药代动力学.世界科学技术-中药现代化,2002,4(3):53-55
72.姜廷良,霍海如.重视中药多组分整合作用的研究.世界科学技术-中药现代化,2002,5(2):1-3
73.王升启.中药基因组学与中药蛋白质组学.世界科学技术-中药现代化,2000;2(1)4-8
74.中药现代化科技发展战略研究课题组.中药现代化重大基础理论研究与重点任务.世界科学技术-中药现代化,2001;3(6):1-6
75.王忠,王阶,王永炎.后基因组时代中医证候组学研究的思考.中国中西医结合杂志,2001;8(21):621-623
1.Devaux PG,Horning MG,Horning EC.Benyzl-oxime derivative of steroids:a new metabolicprofile procedure foe human urinary steroids.Anal Lett,1971,4:151）
2.FiehnO．Metabolic networks of Cucurbita maxima phloem．Phytochemistry．2003，62(6)：875
3.FiehnO．Link between genotypes and phenotypes．Plant M ol Biol，2002，48(2)：155
4.Taylor J，King R D，Altmann T，et al．Application of metabolomics to plant genotypediscrimination using statistics and machine learning．Bioinformatics，2002，18(3)：241
5.Hall R，Beale M，Fiehn O，et a1．Plant metabolomics：the missing link in functionalgenomics strategies． Plant Cell，2002，14(7)：1437
6. Nicholson J K，Lindon J C，Holmes E． Metabonomics ：understanding the metabolicresponsesof living systems to pathophysi-ological stimuli via muhi-variate statisticalanalysis of biological NMR spectroscopic data．Xenobiotica.1999，29(11)：1181
7.刘昌孝.代谢组学的发展于药物研究开发.天津药学.2005，17（2）：1－6）
8.Gennan JB，Bauman DE，Burrrin DG，et al.Metabolomics in the opening decade of the 21stcentury:building the roads to individualized health．J Nutn，2004，134(10)：2729
9Nicholson J K,Holmes E,Wilson I D.Gut microorganisms,mammalian metabolism andpersonalized health care.Nat Rev Microbiol,2005,3:431~438.
10Xu J,Gordon J I.Inaugural Article:Honor thy symbionts.Proc.Natl.Acad.Sci.USA,2003,100:10452-10459.
11．朱航，唐惠儒，张许，刘买利.基于NMR的代谢组学研究.化学通报.2006,69(7):1-9
12.赵剑宇,颜贤忠.基于核磁共振的代谢组学研究进展.国外医学药学分册，2004，31（5）：308～312
13.徐旻，林东海，刘昌孝.代谢组学研究现状与展望.药学学报，2005，40（9）：769～774
14.Nicholson J K，Wilson I D．High resolution proton magnetic resonance spectroscopy ofbiological fluids．Prog Nucl M ag Res Spectrosc，1989，21(4)：449
15.Tang H，Wang Y，Nicholson J K，et a1．Use of relaxation-edited one-dimensional and twodimensional nuclear magnetic resonance spectroscopy to improve detection of smallmetabolites in blood plasma．Anal Biochem,2004，325(3)：260
16.Backwith Hall BM。Thompson N A，Nicholson J K，et a1．A metabonomic investigationof hepatotoxicity using diffusion edited 1H NMR spectroscopy of blood serum．Analyst。2003，128(7)：814
17. Griffin J L, Cemal C K, Pook M A. Defining a metabolic phenotype in the brain of a transgenic mouse model of spinocerebeUar ataxia3. Physiol Genomics, 2004, 16(3): 334
18.Gamache PH, Meyer DF, Granger MC, et al. Metabolomic applications of electrochemistry/mass spectrometry. J Am Soc Mass Spectrom, 2004, 15:1717
19.陈慧梅.代谢组学及其研究方法和应用.肾脏病与透析肾移植杂志，2005，14（1）：59～63
20. Smith LL. Key challenges for toxicologists in the 21st Century. Trends Pharmacol Sci. , 2001, 22(6): 281 285
21. Gavaghan CL, Holmes E, Lenz E, etal. An NMR-based metabonomic approach to investigate the biochemical consequences of genetic strain differences: application to the C57BL10J and Alpk: ApfCD mouse. FEBS Lett, 2000, 484(3): 169 174
22. Jones AR. Some observations on the urinary exeretion of glyeine conjugates by laboratory animals. Xenobiotica 1982, 12: 387
23. Bollard ME, Keun H, Ebbels T. et al. Comparative metabonomics of diferential species toxicity of hydrazine in the rat and mouse. Toxicol, 2005, in press
24. Costa C, DeAntoni A. Baccichetti F. etal. Metabolites and enzyme activities involved in tryptophan metabolism in two strains of mouse. Ital J Biochem, 1984, 33(5): 319
25. Stanley EG. 'H NMR spectroscopic and chemometrie studies on endogenous physiological variation in rats. Ph D thesis, University of London, 2002, 43
26. Rikans LE, Hombrook KR. Cai Y. Carbon tetrachloride hepatotoxicity as a function of age in female Fischer 344 rats. Mech. Ageing Dev, 1994, 76(2-3): 89
27. KishikawaT, Takahashi H, Shimaza aE, etal. Diurnal changes in calcium and phosphate metabolism in rats. Horm Metab Res.1980, 12: 545
28. Phipps AN, Stewart J, Wright B, et al. Effect of diet on the urinary excretion of hippuric acid and other dietary-derived aromatics in rat. A complex interaction between diet gut microflora and substrate specificity. Xenobiotica. 1998, 28(5): 527
28. Honglian S, Vigneau-Callahan KE, Shestopalov Al, et al. Characterisation of diet-dependent metabolic serotypes: primary validation of male and female serotypes in independent cohorts of rats. J Nutr. 2002, 132: 1039
29. Zuppi C, Messana 1, Forni F, et al. 'H NMR spectra of normal urines: reference ranges of the major metabolites. Clin Chim Acta, 1997, 265(1): 85
30. Lindon JC. Nicholson JK. Everett JR. NMR spectroscopy of biofluids. Annum Reports on NMR Spectroscopy, 1999, 38:1
31. M A Constantinou, E Papakonstantinou, M Spraul et al. H NMR-based metabonomics for the diagnosis of inborn errors of metabolism in urine . Anal. Chim. Acta, 2005, 542: 169-177
32. M A Constantinou, E Papakonstantinou, D Benaki et al. Application of nuclear magnetic resonance spectroscopy combined with principal component analysis in detecting inborn errors of metabolism using blood spots: a metabonomic approach .Anal. Chim. Acta, 2004,511: 303-312
33. SHMoolenaar, UFHEngelke, RA Wevers. Proton nuclear magnetic resonance spectroscopy of body fluids in the field of inborn errors of metabolism. Ann. Clin. Biochem., 2003, 40: 16-24
34. J T Brindle, H Antti, E Holmes et al. Rapid and noninvasrve diagnosis of the presence and severity of coronary heart disease using 'H-NMR-based metabonomics. Nat. Med., 2002, 8:1439-1444
35. J T Brindle, J K Nicholson, P M Schofield et al. Application of chemometrics to 'H NMR spectroscopic data to investigate a relationship between human serum metabolic profiles and hypertension. Analyst, 2003, 128: 32-36
36. M A Burns, W L He, C L Wu et al. Quantitative Pathology in Tissue MR Spectroscopy Based Human Prostate Metabolomics Technol. Cancer Res. Treat, 2004, 3: 591-598
37. K Odunsi, R Wollman, C Ambrosone et al. Early Detection and Diagnosis :Detection of epithelial ovarian cancer using 1H NMR-based metabonomics. J. Soc. Gynecol. Invest. 2004, 11: 70A-70A
38. Y L Wang, E Holmes, JK Nicholson, et al. Proc. Natl. Acad. Sci. U. S. A., 2004
39. J Yang, G W Xu, Q F Hong et al. Discrimination of Type 2 diabetic patients from healthy controls by using metabonomics method based on their serum fatty acid profiles. J. Chromatogr. B, 2004, 813: 53-58
40. C Wang, H W Kong, Y F Guan et al. Plasma Phospholipid Metabolic Profiling and Biomarkers of Type 2 Diabetes Mellitus Based on High-Performance Liquid Chromatography/Electrospray Mass Spectrometry and Multivariate Statistical Analysis .Anal. Chem., 2005, 77: 4108-4116
41. H R Tang, Y L Wang, J C Lindon et al. UKP: 0323451.5
42. SlimRM, Roberstson D G, AjbassamM, et al. Effict od dexamethasone on the metabonomics profile associated with phosphodiesterase inhibitor-induced vascular lesions in rats. Toxicol ApplRas, 2003, 183(2): 108
43. Watkins S M , Reifsnyder P R, Pan H J. et al. lipid metabolomewide effects of the PPAR gamma agonist rosiglitazone. Lipid res,2002, 43 (11): 1809
44. Zhang Z P, Su Y, Yue N, Zhao Y G. Bichemical mechanism of comparesion studies of clozapine and quetiapine by using metabonomic method in rats. Asian J. Drug Metabolism and Pharmacokinetics, 2004, 4 (4): 281
45. J C Lindon, J K Nicholson, E Holmes et al. Contemporary issues in toxicology the role of metabonomics in toxicology and its evaluation by the COMET project.Toxicol. Appl. Pharmacol, 2003, 187: 137-146
46.J C Lindon, H C Keun, T M D Ebbels et al. The Consortium for Metabonomic Toxicology (COMET): aims, activities and achievements. Pharmacogenomics, 2005,6: 691-699
47. Geisow MJ. Proteomics: one small step for a digital computer, one giant leap for humankind. Not Biotechnol.1998, 16(2): 206
48.Eisen MB，Spellman PT，Brown PO，et a1．Ouster analysis and display of genome-wideexpression patterns．Proc Natl Acad Sci USA，1998，95(25)：14863 14868
49. Holmes E，NiehoUs AW，Lindon JC，et a1．Chemometric models for toxicity classification based on NMR spectra ofbiofluids．ChemResToxicol.2OOO，13(6)：471-478
50.Raamsdonk LM，Teusink B，Broadhurst D，et al．A functional genomics strategy that usesme tabolome data to reveal the phenotype of silent mutations．Nat Biotechnol.2001，19(1)：45 50
51.Bandy JG，Spurgeon DJ，Svendsen C．et al．Earthworm species of the genus Eisenia canbe phenotypically differentiated by metabolic profiling．FEBS Lett.2OO2，521(1-3)：115-120．
52.KS Solanky,N J Bailey,B M Beckwith-Hall.et al.Biofluid 1H NMR-based metabonomictechniques in nutrition research metabolic effects of dietary isoflavones in humans.Nutr.Biochem.,2005,16:236-244.
53.YL Wang,HR Tang,JK Nicholson et al.A metabonomic strategy for the detection of themetabolic effects of chamomile(MatricariarecutitaL.)ingestion.Agric FoodChem.2005,53:191-196.
54.贾伟，蒋健，刘平等.代谢组学在中医药复杂理论体系研究中的应用.中国中药杂志，2006，31（8）：621～624.
55.王米渠,张卫,李珉,等.基因芯片筛选针刺衰老鼠涌泉穴的318个差异表达基因初报.中国中西医结合杂志,2002,22(11):928.
56.王米渠,冯韧,严石林等.基因表达谱芯片发现热药疗寒的相关七类基因初报.中医杂志，2003,44(4):228.
57.王建红,罗来成,于润桥.基于系统工程理论的中医方证研究.辽宁中医杂志.2006;33(3):296
58.王文健,沈自尹,张新民,等.补肾法对老年男性下丘脑-垂体-性腺轴作用的临床和实验研究.中医杂志1986;27(4):32
59.蔡定芳,沈自尹,张玲绢,等.右归饮对大鼠下丘脑-垂体-肾上腺-胸腺轴功能抑制模型的影响.中国免疫学杂志1994;10(7):236
60.蔡定芳,沈自尹,陈晓红,等.乌头碱对大鼠下丘脑促肾上腺皮质激素释放激素含量的影响.中国中西医结合杂志1996;16(9):544
61.钟历勇,沈自尹,蔡定芳,等.补肾健脾活血三类复方对下丘脑-垂体-肾上腺-胸腺轴及CRF基因表达的影响.中国中西医结合杂志1997;17(1):39
62.俞瑾,陈红英,毛秋芝.肾主生殖的实验研究.中西医结合杂志1989;9(9):548
63.俞瑾.肾主生殖与生命网络研究中的启示.中国中西医结合杂志2000;20(6):409
64.沈一丁，费贵强，张宇.代谢组学促进中医药现代化.精细化工.2005，22（6）：444-446
65.许国旺，杨军．代谢组学及其研究进展．色谱，2003，21(4)：316 320．p74-76
1.陈主初.病理生理学第 1版.北京:人民卫生出版社.2006:122-139
2.周爱儒.生物化学第 6版.北京:人民卫生出版社.2006:202-347
3.李天心主编,医学心理学,北京:北京医科大学协和医科大学联合出版社,1998:148-159。
4.BornsteinSR,ChrousosGP.Adrenocorticotropin(ACTH) andN0n-ACTH MediatedRegulationoftheAdrenalCortex:NeuralandImmuneInputs.ClinicEndocrinolMetabolism,1999,84(5):1729-1736.
5.亓晓丽,姚树桥.应激与海马可塑性及其机制的研究进展.中国行为医学科学.2003;11(3):356-358
6.McEwen BS. Stress and hippocampal plasticity.Annu Rev Neurosci.1999;22:105-122。
7.张艳美,杨权,许崇涛,等.慢性应激对大鼠海马锥体细胞形态结构的效应.生物化学与生物物理进展.2002;29(5):719-723
8.McEwenBS.Effectsofadverseexperiencesforbrainstructureandfunction.BiolPsychiatry.2000,48(8):721-731
9.VyasA.Chronicstress inducescontrastingpatternsofdendriticremodelinginhippocampalandamygdaloidneurons.JNeurosci.2002,22(15):6810-6818
10.敖海清,徐志伟,严 灿,等.逍遥散对应激大鼠海马突触体内PKC活性及Ca2+浓度的影响.山东中医杂志.2006,25(2):112-114
11.唐已婷,陈家旭.束缚应激与中医肝的关系.中国医药学报.2002;17(2):82-83
12.唐已婷,陈家旭.慢性束缚应激的中枢介质-促皮质释放激素[J].中国行为医学科学,2001,10(4):394-396
13.唐已婷,陈家旭.慢性束缚应激动物模型的研究进展(综述)[J].北京中医药大学学报,2002,25(1):225-28
14.唐已婷,陈家旭.三种中药复方对慢性束缚应激大鼠下丘脑-垂体-肾上腺轴的调节[J].北京中医药大学学报,2002,25(3):23-26
15.赵歆,陈家旭,杨建新,等.疏肝中药复方对慢性束缚应激大鼠下丘脑 垂体 肾上腺轴的调节[J].实验动物科学与管理,2003,20(2):6-12
16.陈家旭,唐已婷.慢性束缚应激模型大鼠相关脑区前阿黑皮素基因表达的变化及中药复方的影响[J].中国药物与临床,2003,3(3):223-226
17.李伟,陈家旭,杨建新,等.疏肝、健脾、补肾复方对慢性束缚应激大鼠行为学和免疫功能的影响[J].中国实验动物学报,2003,11(1):33-37
18.陈家旭,唐已婷.逍遥散对慢性束缚应激模型大鼠相关脑区 CRF基因表达的影响[J].中国应用生理学杂志,2004,20(1):71-74
19.陈家旭,杨建新,赵歆,等.慢性束缚应激大鼠下丘脑β-内啡肽变化及中药复方对其的影响[J].中国医药学报,2004,19(2):83-85
20.陈家旭,李伟,赵歆,等.三种中药复方对慢性束缚应激大鼠行为及皮层和海马 NT3的影响[J].北京中医药大学学报,2004,27(2):19-23
21.马梦茵,陈家旭.论辨证论治的发展方向[J].中国医药学报,2004,19(6):367-369
22.ChenJX,LiW,ZhaoX,YangJX,XuHY,WangZF,YueGX.ChangesofmRNAexpressionofenkephalinandprodynorphininhippocampusofratswithchronicimmobilizationstress.WorldJGastroenterol.2004;10(17):2547-9
23.岳广欣,陈家旭,王竹风.肝主疏泄的生理学基础探讨[J].北京中医药大学学报,2005,02:1-4.
24.陈家旭,李伟,赵歆,等.慢性束缚应激大鼠海马脑啡肽mRNA和前强啡肽mRNA表达及中药复方的影响[J].中国应用生理学杂志,2005,21(2):121-125
25.王竹风,陈家旭,岳广欣,等.刍议肝郁理论及其影响[J].中华中医药杂志,2005,20(6):367-368
26.岳广欣,陈家旭,王竹风.中医心肝肾三脏在应激反应中的作用 [J].辽宁中医杂志,2005,32(6):528-530
27.李伟,陈家旭.慢性束缚应激大鼠海马 BDNFTrkBNT_3的变化及逍遥散对其影响[J].中医药学刊,2005,23(7):1205-1208
28.王竹风,陈家旭,岳广欣.慢性应激与海马神经元的关系探析[J].辽宁中医学院学报,2005,7(4):337-339
29.王 竹 风 ,陈 家 旭 ,赵 歆 ,等 .肝 主 升 发 在 五 脏 中 的 作 用 探 微 [J].辽 宁 中 医 杂 志 ,2005,32(8):776-777
30.王竹风,陈家旭,赵歆,等.讨肝郁的影响[J].时珍国医国药,2005,16(10):948-949
31.徐洪雁,赵歆,陈家旭,等.慢性束缚应激大鼠海马 CA1区 L-ENK的变化及逍遥散的调节作用.世界科学技术-中医药现代化 2005;7(5):18-22
32.王竹风.慢性束缚应激对脑区 NMDA受体及生长相关蛋白的影响和逍遥散的调节作用.北京中医药大学博士研究生学位论文.2006:3
33.岳广欣. 束缚应激大鼠中枢AMPA受体及相关蛋白变化与肝主疏泄的关系.北京中医药大学博士研究生学位论文.2006:130
34.陈家旭,杨维益.神经-内分泌-免疫网络研究概况及其与中医肝脏关系的探讨.北京中医药大学学报.1995;18(4):7-11
35.严灿,徐志伟,李艳,等.调肝、补肾、健脾方药对慢性心理应激大鼠单胺类神经递质影响的比较研究.中国中西医结合杂志.2002;22(12):925-928
36.徐志伟,严灿,李艳,等.慢性心理应激大鼠血浆及中枢β-EP、ACTH含量的变化及调肝治法方药的影响.中药药理与临床.2002;18(5):37-39
37.王朝勋,郑洪新,王继伟,等.怒伤肝与神经 内分泌 免疫系统失调探析.辽宁中医杂志1997;24(5)∶205-206
38.严灿,邓中炎,王剑.调肝方药对束缚应激大鼠神经内分泌免疫功能的调节作用.中国病理生理杂志,2000,16(6):560-562
39.叶晓平,宋纯清.柴胡皂苷药理研究进展.中草药,2004,35(12):1434-1436
40.唐伟军,卢新华,周大现,等.柴胡镇痛效应的研究.湖南中医杂志,2003,19(4):52-53
41.李芳,李建北,张东明.柴胡的药理研究进展.时珍国医国药,2004,15(2):120-121
42.史青,聂淑琴,黄璐琦.柴胡属植物化学成分及药理研究新进展.中国实验方剂学杂志,2002,8(5):53-56
43.刘琳娜,梅其炳,程建峰.当归挥发油研究的进展.中成药,2002,24(8):621-623
44.夏泉,张平,李绍平,等.当归的药理作用研究进展.时珍国医国药,2004,15(3):164-166
45.张艳,明亮,王瑜,等.白芍总甙的抗惊厥作用.中国药理学通报,1994,10(5):372-374.
46.张 鸿 炼 , 吴 文 蔚 . 白 芍 总 甙 的 药 理 作 用 与 临 床 应 用 . 中 国 医 院 药 学 杂 志 ,1995,15(8):371-372
47.周辉,张安平,陈敏珠,等.白芍总甙对应激大鼠下丘脑-垂体-肾上腺轴和免疫功能的调节.中国药理学通报,1994,10(6):429-432
48.邓 兆 智 , 钟 秋 生 . 白 芍 总 苷 的 药 理 与 临 床 研 究 考 释 . 实 用 中 医 内 科 杂 志 ,2004,18(3):181-182
49.邓兆智,钟秋生,刘文国,等.芍药甙对体外培养的大鼠海马和隔区神经元的影响.解剖学研究,2004,24(4):270-272
50.杨翠平,劳业兴,吴凤薇,等.白术的研究进展.中药材,2002,25(3):206-208.
52.徐丽珊,金晓玲,邵邻相.白术及白术多糖对小鼠学习记忆和抗氧化作用的影响.科技通报,2003,19(6):513-515
52.国家中医药管理局《中华本草》编委会.中华本草.第 1版.上海:上海科学技术出版社,1999
53.李素民,杨秀岭,赵智,等.干姜和生姜药理研究进展.中草药,1999,30(6):471-473.
54.梁呈元,李维林,张涵庆,等.薄荷化学成分及其药理作用研究进展.中国野生植物资源,2003,22(3):9-12
1.孟凡胜,隗学玲,沈烈行 .逍遥散药理研究及临床应用进展.国医论坛,2005,20(3):52-53
2.马增春,高月,谭洪玲,梁乾德,王升启.用分子中药组学技术研究四物汤补血的作用机理.世界科学技术中医药现代化,2005,7(3):24-28
3.刘昌孝.中药药代动力学研究的难点和热点. 药学学报, 2005,40(5):395-401
4.李峰,康廷国.系统论与中药复方作用机制探讨.中医药学刊,2003,21(9):1479-1480
5.王本祥,周秋丽.关于中药活性成分的认识及其研究方法.中国中药杂志,2001,26(1):10-13
6.潘卫松,刘美风,石钺,等.血清药理学、血清化学和中药药代动力学.世界科学技术-中药现代化,2002,4(3):53-55
7.姜廷良,霍海如.重视中药多组分整合作用的研究.世界科学技术-中药现代化.2002,5(2):1-3
8.王升启.中药基因组学与中药蛋白质组学.世界科学技术-中药现代化,2000,2(1):4-8
9.Gennan JB，Bauman DE，Burrrin DG，et al.Metabolomics in the opening decade of the 21stcentury:building ther oads to individualized health．J Nutn，2004，134(10)：2729
10.中药现代化科技发展战略研究课题组.中药现代化重大基础理论研究与重点任务.世界科学技术-中药现代化,2001;3(6):1-6
11.王忠,王阶,王永炎.后基因组时代中医证候组学研究的思考.中国中西医结合杂志,2001;8(21):621-623
12.颜贤忠,赵剑宇,彭双清,廖明阳.代谢组学在后基因组时代的作用.波谱学杂志,2004,21(2):263-271
13.Holmes E,Nicholls AW,Lindon JC,Connor SC,Connelly JC,Haselden JN,Damment SJ,Spraul M,Neidig P,Nicholson JK.Chemometric models for toxicity classification basedon NMR spectra of biofluids.Chem Res Toxicol.2000 Jun;13(6):471-8.
14.Waters NJ,Waterfield C J,Farrant R D,et al.Metabonomic Deconvolution Of EmbeddedToxicity:Application To Thioacetamide Hepato-and Nephrotoxicity.Chem Res Toxicol.2005,18(4):639-654.
15.Lindon JC,Nicholson J K,Holmes E,et al.Metabonomics:Metabolic processes studiedby NMR spectroscopy of biofluids.Concepts Magn Reson.2000,12:289-320.
16.王竹风.慢性束缚应激对脑区 NMDA受体及生长相关蛋白的影响和逍遥散的调节作用.北京中医药大学博士研究生学位论文.2006:3
17.岳广欣. 束缚应激大鼠中枢 AMPA受体及相关蛋白变化与肝主疏泄的关系.北京中医药大学博士研究生学位论文.2006:130
18.石慧琳,程义勇,李树田等.慢性束缚应激对大鼠肠粘膜屏障功能的影响.中国行为医学科学.2003,12(3):251-253
19.吕志平,刘承才. 肝郁 大鼠的脂质过氧化反应及逍遥散的保护作用.山东中医学院学报,1995,19(3):199
20.吕志平,刘承才. 肝郁 大鼠血浆TXA2、PGI2水平与肝微循环变化及逍遥散作用.中国微循环,2000,4(3):160
21.唐已婷,陈家旭.束缚应激与中医肝的关系.中国医药学报.2002;17(2):82-83
22.唐 已 婷 ,陈 家 旭 .慢 性 束 缚 应 激 的 中 枢 介 质 -促 皮 质 释 放 激 素 .中 国 行 为 医 学 科学,2001,10(4):394-396
23.唐 已 婷 ,陈 家 旭 .慢 性 束 缚 应 激 动 物 模 型 的 研 究 进 展 (综 述 ).北 京 中 医 药 大 学 学报,2002,25(1):225-28
24.唐已婷,陈家旭.三种中药复方对慢性束缚应激大鼠下丘脑-垂体-肾上腺轴的调节.北京中医药大学学报,2002,25(3):23-26
25.赵歆,陈家旭,杨建新,李伟,刘晓兰,康纯洁,刘燕.疏肝中药复方对慢性束缚应激大鼠下丘脑垂体 肾上腺轴的调节.实验动物科学与管理,2003,20(2):6-12
25.陈家旭,唐已婷.慢性束缚应激模型大鼠相关脑区前阿黑皮素基因表达的变化及中药复方的影响.中国药物与临床,2003,3(3):223-226
27.李伟,陈家旭,杨建新,赵歆,唐已婷,刘晓兰,徐洪雁.疏肝、健脾、补肾复方对慢性束缚应激大鼠行为学和免疫功能的影响.中国实验动物学报,2003,11(1):33-37
28.陈家旭,唐已婷.逍遥散对慢性束缚应激模型大鼠相关脑区 CRF基因表达的影响.中国应用生理学杂志,2004,20(1):71-74
29.陈家旭,杨建新,赵歆,李伟,陈青红,马梦茵,李柳骥,汪南玥,薛飞飞,洪梅.慢性束缚应激大鼠下丘脑β-内啡肽变化及中药复方对其的影响.中国医药学报,2004,19(2):83-85
30.陈家旭,李伟,赵歆,杨建新,陈青红,马梦茵,李柳骥,徐洪雁.三种中药复方对慢性束缚应激大鼠行为及皮层和海马 NT3的影响.北京中医药大学学报,2004,27(2):19-23
31.马梦茵,陈家旭.论辨证论治的发展方向.中国医药学报,2004,19(6):367-369
32.Chen JX,Li W,ZhaoX ,etall.Changes of mRNA expression of enkephalin and prodynorphin in hippocampus of rats with chronic immobilization stress.World J Gastroenterol.2004;10(17):2547-9
33.岳广欣,陈家旭,王竹风.肝主疏泄的生理学基础探讨.北京中医药大学学报,2005,02:1-4
34.陈家旭,李伟,赵歆,杨建新,徐洪雁.慢性束缚应激大鼠海马脑啡肽mRNA和前强啡肽mRNA表达及中药复方的影响.中国应用生理学杂志,2005,21(2):121-125
35.王竹风,陈家旭,岳广欣等.刍议肝郁理论及其影响.中华中医药杂志.2005,20(6):367-368
36.岳广欣,陈家旭,王竹风.中医心肝肾三脏在应激反应中的作用.辽宁中医杂志.2005,32(6):528-530
37.李伟,陈家旭.慢性束缚应激大鼠海马 BDNFTrkBNT_3的变化及逍遥散对其影响.中医药学刊,2005,23(7):1205-1208
38.王 竹 风 ,陈 家 旭 ,岳 广 欣 .慢 性 应 激 与 海 马 神 经 元 的 关 系 探 析 .辽 宁 中 医 学 院 学报,2005,7(4):337-339
39.王竹风,陈家旭,赵歆,岳广欣,汪宝军.肝主升发在五脏中的作用探微.辽宁中医杂志,2005,32(8):776-777
40.王竹风,陈家旭,赵歆,岳广欣,汪宝军.讨肝郁的影响.时珍国医国药,2005,16(10):948-949
41.徐洪雁,赵歆,陈家旭,马梦茵,王竹风,岳广欣.慢性束缚应激大鼠海马 CA1区 L-ENK的变化及逍遥散的调节作用.世界科学技术-中医药现代化 2005;7(5):18-22
42.张艳美.慢性应激、大脑损害与抑郁症.国外医学-精神病分册.2001,28(2):105-109
43.王良斌,张维宁.大鼠不同脑区突触体钙水平的年龄差异.动物学研究,1995,16(4):373-377
44.耿战辉,程义勇,洪燕等.心理应激时大鼠海马神经元的钙状态变化及锌的作用研究.营养学报,2003,5(1):65-68
45.牛玉杰,石年.溴氰菊酯对大鼠神经细胞内游离钙的影响.卫生毒理学杂志,2001,15(4):216-218
46.William M.Vasopressin-induced protein kinase C-dependent superoxide generation contributes to ATP-seneitive potassium channel butnot calcium-sensitive potassium channelfunction impairment after brain injury.Stroke,2001,32(6):1408-1414
47.敖海清,徐志伟,严灿等.逍遥散对应激大鼠海马突触体内PKC活性及Ca2+浓度的影响，山东中医杂志.2006,25（2）:112-114
48.MacManus J P,Hill IE,Preston E,et al.Difference in DNA fragmentation followingTransient cerebral or decapitation ischemia in rat.J Cereb Blood Flow Metab,1995,15(5):728-737
2.王阶、张兰凤、王永炎.方证对应理论源流及临床研究.世界科学技术-中医药现代化.2004,6(4):13-17
3.李致重.日本汉方医学衰落轨迹.崔月犁.中医沉思录.北京:中医古籍出版社,1997,162-170.
4.刘渡舟.方证相对论.北京中医药大学学报.1996,19(1):3
5.谢鸣. 方证相关 逻辑命题及其意义.北京中医药大学学报,2003,26(2):11-12
6.李瑞,尹英杰,林殷,鲁兆麟.中医方证客观化研究初探.北京中医药大学学报 2002;25(3):6-10
7.窦志芳,郭 蕾,张俊龙等. 方证对应 研究的思考.中医杂志.2006,47(8):563
8.王阶.方证对应与方证标准规范探讨.中医杂志 ,2002;43(7):489
9.俞雪如.柯琴 以方名证 思想对吉益东洞 方证相对说 的影响.上海中医药大学学报,2001,15(2):2
10.李国臣,王冠民,崔文艺.胡希恕方证辨证说略.上海中医药杂志,2003,37(10):39.
11.邓铁涛.再论辨证论治.新中医,1999,31(4):8
12.金海浩.中医学的方证相应论.黑龙江中医药,2005,1(1):45
13.张兰凤,王 阶,王永炎.方证对应研究.中华中医药杂志,2005,20(1):8
14.刘晋平,黄宇虹,陆小左.数据挖掘在中医脉诊中的应用.天津中医学院学报.2003,22(3):9
15.姚美村,袁月梅,艾路.数据挖掘及其在中医药现代化中的应用.北京中医药大学学报,2002,25(5):20
16.刘娟,蒋永光,胡波等.白术类方的药证关联分析.成都中医药大学学报.2004,27(4):5
17.孙占全,刘艳骄,西广成等.证候与方剂相关性的研究.中国中医基础医学杂志.2006,12(2):150
18.王米渠,张卫,李珉,等.基因芯片筛选针刺衰老鼠涌泉穴的 318个差异表达基因初报 .中国中西医结合杂志,2002,22(11):928.
19.王米渠,冯韧,严石林等.基因表达谱芯片发现热药疗寒的相关七类基因初报.中医杂志,2003,44(4):228
20.高月,刘永学,马增春等.白芍在四物汤中的反佐作用.中国实验方剂学杂志,2004,10(2):26-28
21.郭平,马增春,李鹰飞等.四物汤对放射线致血虚证小鼠骨髓蛋白质表达的影响.中国中药杂志,2004,29(9):893-896
22.梁乾德,路晓钦,马增春等.四物汤促进造血功能成分的初步研究.中国中药杂志,2004,6:546-549
23.马增春,高月,谭洪玲等.四物汤对辐射致血虚证小鼠血清蛋白质表达的影响.中国中药杂志,2003,28(11):1050-1054
24.高月,马增春,刘永学等.四物汤及其提取物对辐射致血虚证小鼠造血作用的研究.天津中医药,2003,6:47-51
25.高月,马增春,刘永学等.CD34+细胞在血虚证小鼠中的变化及意义.中华中西医杂志,2003,4(5):658-659
26.佟丽,陈苏红,马增春等.用寡核苷酸芯片研究血虚小鼠细胞因子基因表达谱.中草药,2003,7:625-629
27.高月,马增春,陈鹏等.四物汤及其各单药对辐射小鼠骨髓造血的调节作用.中药新药与临床药理,2002,12(5):336-339
28.马增春,高月,谭洪玲等.实验性血虚证小鼠骨髓损伤机制的研究.中医杂志,2002,43(增刊):73-74
29.谭洪玲,马增春,高月等.四物汤对血虚证小鼠骨髓细胞CD34抗原表达的影响.中药新药与临床药理,2002,13(1):11-13
30.马淑华,王升启,范保星等.抑制消减杂交和辐射小鼠血虚模型筛选 四物汤 诱导表达的基因.生物技术通讯,2002,13(2):214-217
31.马增春,高月,刘永学等.辐射小鼠骨髓中CD34+细胞的变化及其意义.中华放射医学与防护杂志,2001,21(1):12-15
32.马增春,高月,刘永学等.环磷酰胺所致血虚证小鼠骨髓CD34+细胞的变化及其意义.中国基础医学杂志,2001,7(7):42-44
33.马增春,高月,刘永学等.四物汤对环磷酰胺所致血虚证小鼠造血细胞作用的研究.中国实验方剂学杂志,2001,7(5):13-15
34.马增春,高月,刘永学等.四物汤对 γ 射线照射致血虚证小鼠造血细胞作用的研究.中国实验方剂学杂志,2001,7(3):41-44
35.谭洪玲,高月,马增春等.四物汤刺激 Peyer sPatch促进骨髓造血的研究.中药药理与临床,2001,17(2):1-3
36.路晓钦,马增春,高月.四物汤药理研究进展.中国实验方剂学杂志,2001,7(4):56-69
37.马增春,高月.血虚证的病理研究概况.新中医,2000,32(9):60-61
38.马增春,高月,刘永学等.再生障碍性贫血患者骨髓造血调控机制的研究进展.国外医学输血及血液学分册,2000,5:351-354
39.刘永学,马增春,高月等.实验性血虚证小鼠骨髓造血干/祖细胞的变化.中国基础医学杂志,2000,6(增刊):337-341
40.董波,马增春,高月等.碘化丙啶对小鼠骨髓 CD34细胞计数的影响.中国应用生理学杂志,2000,16(3):133
41.廖建群,钱锋,马增春等.中医补血方剂对受照小鼠造血功能的影响.生物技术通讯,2000,11(3):214
42.Gennan JB,Bauman DE,Burrrin DG,et al.Metabolomics in the opening decade of the21st century:building the roads to individualized health.J Nutn,2004,134(10):2729
43.GavaghanCL,Holmes E,I_enz E,et al .AnNMR-based metabonomic approach to investigate the biochemi calconsequences of genetic strain diferencesi application to the C57BL10J and AIpk:ApfCDmouse[J].FEBSLett,2O00,484(3):169 174
44.赵剑宇,颜贤忠.基于核磁共振的代谢组学研究进展.国外医学药学分册,2004,31(5):308～
31245.沈一丁,费贵强,张 宇.代谢组学促进中医药现代化.精细化工,2005,22(6):444
46.贾 伟,蒋 健,刘 平等.代谢组学在中医药复杂理论体系研究中的应用.中国中药杂志.2006,31(8):621
47.王桐生,谢鸣.代谢组学与中医药现代化.中医杂志.2006,47(10):723-725
48.衷敬柏,王阶,赵宜军等.病证结合与方证相应研究.辽宁中医杂志.2006;33(2):137-139
49.李茹柳,陈艳芬,陈蔚文.左金丸方证相应动物模型建立的回顾及其对方证研究的意义.广州中医药大学学报.2005;22(3):236-238
50.陈艳芬,陈蔚文,李茹柳.左金丸与反左金丸的方证对应实验研究.广东药学院学报2004;20(5):501-503
51.年莉,方玲,刘鸿雁.吴茱萸汤方证学研究.天津中医药.2004;21(1):38-40
52.费忠东.大柴胡汤现代方证研究.江苏中医药.2003;24(8):43-45
53.王阶,荆鲁,衷敬柏.血府逐瘀汤拆方临床研究.中国中药杂志.2004;29(8):803-807
54.张兰凤,王阶,衷敬柏,等.冠心病病证结合方证对应临床研究.中医杂志.2004;45(6):444
55.张杰.利用方证相应学说探寻中医证的物质基础.中国中医药信息杂志.2005;12(11):3-5
56.李沛清,刘喜平.对中医证与方证研究的思考.中国中医药信息杂志.2005;12(4):5-6
57.王建红,罗来成,于润桥.基于系统工程理论的中医方证研究.辽宁中医杂志.2006;33(3):296
58.王文健,沈自尹,张新民,等.补肾法对老年男性下丘脑-垂体-性腺轴作用的临床和实验研究.中医杂志 1986;27(4):32
59.蔡定芳,沈自尹,张玲绢,等.右归饮对大鼠下丘脑-垂体-肾上腺-胸腺轴功能抑制模型的影响.中国免疫学杂志 1994;10(7):236
60.蔡定芳,沈自尹,陈晓红,等.乌头碱对大鼠下丘脑促肾上腺皮质激素释放激素含量的影响.中国中西医结合杂志 1996;16(9):544
61.钟历勇,沈自尹,蔡定芳,等.补肾健脾活血三类复方对下丘脑-垂体-肾上腺-胸腺轴及 CRF基因表达的影响.中国中西医结合杂志 1997;17(1):39
62.俞瑾,陈红英,毛秋芝.肾主生殖的实验研究.中西医结合杂志 1989;9(9):548
63.俞瑾.肾主生殖与生命网络研究中的启示.中国中西医结合杂志 2000;20(6):409
64.路晓钦,高月,梁乾德,谭洪玲.四物汤中不同组分对血虚模型小鼠造血功能的影响.中成药2005;7(10):1189-1193
65.陈家旭,唐已婷.逍遥散对慢性束缚应激模型大鼠相关脑区 CRF基因表达的影响.中国应用生理杂志 2004;20(1):71-74
66.陈家旭,杨建新,赵歆,等.慢性束缚应激大鼠下丘脑-内啡肽变化及中药复方作用的影响.中国医药学报 2004;19(2):42-44
67.嵇波,陈家旭,鲁兆麟,等.逍遥散对人体神经内分泌免疫系统的影响.北京中医药大学学报2003;26(6):72-75
68.马增春,高月,谭洪玲,梁乾德,王升启.用分子中药组学技术研究四物汤补血的作用机理.世界科学技术中医药现代化,2005,7(3):24-28
69.刘昌孝.中药药代动力学研究的难点和热点.药学学报,2005,40(5):395-401李峰,康廷国.系统论与中药复方作用机制探讨.中医药学刊,2003,21(9):1479-1480
70.王本祥,周秋丽.关于中药活性成分的认识及其研究方法.中国中药杂志,2001,26(1):10-13
71.潘卫松,刘美风,石钺,等.血清药理学、血清化学和中药药代动力学.世界科学技术-中药现代化,2002,4(3):53-55
72.姜廷良,霍海如.重视中药多组分整合作用的研究.世界科学技术-中药现代化,2002,5(2):1-3
73.王升启.中药基因组学与中药蛋白质组学.世界科学技术-中药现代化,2000;2(1)4-8
74.中药现代化科技发展战略研究课题组.中药现代化重大基础理论研究与重点任务.世界科学技术-中药现代化,2001;3(6):1-6
75.王忠,王阶,王永炎.后基因组时代中医证候组学研究的思考.中国中西医结合杂志,2001;8(21):621-623
1.Devaux PG,Horning MG,Horning EC.Benyzl-oxime derivative of steroids:a new metabolicprofile procedure foe human urinary steroids.Anal Lett,1971,4:151）
2.FiehnO．Metabolic networks of Cucurbita maxima phloem．Phytochemistry．2003，62(6)：875
3.FiehnO．Link between genotypes and phenotypes．Plant M ol Biol，2002，48(2)：155
4.Taylor J，King R D，Altmann T，et al．Application of metabolomics to plant genotypediscrimination using statistics and machine learning．Bioinformatics，2002，18(3)：241
5.Hall R，Beale M，Fiehn O，et a1．Plant metabolomics：the missing link in functionalgenomics strategies． Plant Cell，2002，14(7)：1437
6. Nicholson J K，Lindon J C，Holmes E． Metabonomics ：understanding the metabolicresponsesof living systems to pathophysi-ological stimuli via muhi-variate statisticalanalysis of biological NMR spectroscopic data．Xenobiotica.1999，29(11)：1181
7.刘昌孝.代谢组学的发展于药物研究开发.天津药学.2005，17（2）：1－6）
8.Gennan JB，Bauman DE，Burrrin DG，et al.Metabolomics in the opening decade of the 21stcentury:building the roads to individualized health．J Nutn，2004，134(10)：2729
9Nicholson J K,Holmes E,Wilson I D.Gut microorganisms,mammalian metabolism andpersonalized health care.Nat Rev Microbiol,2005,3:431~438.
10Xu J,Gordon J I.Inaugural Article:Honor thy symbionts.Proc.Natl.Acad.Sci.USA,2003,100:10452-10459.
11．朱航，唐惠儒，张许，刘买利.基于NMR的代谢组学研究.化学通报.2006,69(7):1-9
12.赵剑宇,颜贤忠.基于核磁共振的代谢组学研究进展.国外医学药学分册，2004，31（5）：308～312
13.徐旻，林东海，刘昌孝.代谢组学研究现状与展望.药学学报，2005，40（9）：769～774
14.Nicholson J K，Wilson I D．High resolution proton magnetic resonance spectroscopy ofbiological fluids．Prog Nucl M ag Res Spectrosc，1989，21(4)：449
15.Tang H，Wang Y，Nicholson J K，et a1．Use of relaxation-edited one-dimensional and twodimensional nuclear magnetic resonance spectroscopy to improve detection of smallmetabolites in blood plasma．Anal Biochem,2004，325(3)：260
16.Backwith Hall BM。Thompson N A，Nicholson J K，et a1．A metabonomic investigationof hepatotoxicity using diffusion edited 1H NMR spectroscopy of blood serum．Analyst。2003，128(7)：814
17. Griffin J L, Cemal C K, Pook M A. Defining a metabolic phenotype in the brain of a transgenic mouse model of spinocerebeUar ataxia3. Physiol Genomics, 2004, 16(3): 334
18.Gamache PH, Meyer DF, Granger MC, et al. Metabolomic applications of electrochemistry/mass spectrometry. J Am Soc Mass Spectrom, 2004, 15:1717
19.陈慧梅.代谢组学及其研究方法和应用.肾脏病与透析肾移植杂志，2005，14（1）：59～63
20. Smith LL. Key challenges for toxicologists in the 21st Century. Trends Pharmacol Sci. , 2001, 22(6): 281 285
21. Gavaghan CL, Holmes E, Lenz E, etal. An NMR-based metabonomic approach to investigate the biochemical consequences of genetic strain differences: application to the C57BL10J and Alpk: ApfCD mouse. FEBS Lett, 2000, 484(3): 169 174
22. Jones AR. Some observations on the urinary exeretion of glyeine conjugates by laboratory animals. Xenobiotica 1982, 12: 387
23. Bollard ME, Keun H, Ebbels T. et al. Comparative metabonomics of diferential species toxicity of hydrazine in the rat and mouse. Toxicol, 2005, in press
24. Costa C, DeAntoni A. Baccichetti F. etal. Metabolites and enzyme activities involved in tryptophan metabolism in two strains of mouse. Ital J Biochem, 1984, 33(5): 319
25. Stanley EG. 'H NMR spectroscopic and chemometrie studies on endogenous physiological variation in rats. Ph D thesis, University of London, 2002, 43
26. Rikans LE, Hombrook KR. Cai Y. Carbon tetrachloride hepatotoxicity as a function of age in female Fischer 344 rats. Mech. Ageing Dev, 1994, 76(2-3): 89
27. KishikawaT, Takahashi H, Shimaza aE, etal. Diurnal changes in calcium and phosphate metabolism in rats. Horm Metab Res.1980, 12: 545
28. Phipps AN, Stewart J, Wright B, et al. Effect of diet on the urinary excretion of hippuric acid and other dietary-derived aromatics in rat. A complex interaction between diet gut microflora and substrate specificity. Xenobiotica. 1998, 28(5): 527
28. Honglian S, Vigneau-Callahan KE, Shestopalov Al, et al. Characterisation of diet-dependent metabolic serotypes: primary validation of male and female serotypes in independent cohorts of rats. J Nutr. 2002, 132: 1039
29. Zuppi C, Messana 1, Forni F, et al. 'H NMR spectra of normal urines: reference ranges of the major metabolites. Clin Chim Acta, 1997, 265(1): 85
30. Lindon JC. Nicholson JK. Everett JR. NMR spectroscopy of biofluids. Annum Reports on NMR Spectroscopy, 1999, 38:1
31. M A Constantinou, E Papakonstantinou, M Spraul et al. H NMR-based metabonomics for the diagnosis of inborn errors of metabolism in urine . Anal. Chim. Acta, 2005, 542: 169-177
32. M A Constantinou, E Papakonstantinou, D Benaki et al. Application of nuclear magnetic resonance spectroscopy combined with principal component analysis in detecting inborn errors of metabolism using blood spots: a metabonomic approach .Anal. Chim. Acta, 2004,511: 303-312
33. SHMoolenaar, UFHEngelke, RA Wevers. Proton nuclear magnetic resonance spectroscopy of body fluids in the field of inborn errors of metabolism. Ann. Clin. Biochem., 2003, 40: 16-24
34. J T Brindle, H Antti, E Holmes et al. Rapid and noninvasrve diagnosis of the presence and severity of coronary heart disease using 'H-NMR-based metabonomics. Nat. Med., 2002, 8:1439-1444
35. J T Brindle, J K Nicholson, P M Schofield et al. Application of chemometrics to 'H NMR spectroscopic data to investigate a relationship between human serum metabolic profiles and hypertension. Analyst, 2003, 128: 32-36
36. M A Burns, W L He, C L Wu et al. Quantitative Pathology in Tissue MR Spectroscopy Based Human Prostate Metabolomics Technol. Cancer Res. Treat, 2004, 3: 591-598
37. K Odunsi, R Wollman, C Ambrosone et al. Early Detection and Diagnosis :Detection of epithelial ovarian cancer using 1H NMR-based metabonomics. J. Soc. Gynecol. Invest. 2004, 11: 70A-70A
38. Y L Wang, E Holmes, JK Nicholson, et al. Proc. Natl. Acad. Sci. U. S. A., 2004
39. J Yang, G W Xu, Q F Hong et al. Discrimination of Type 2 diabetic patients from healthy controls by using metabonomics method based on their serum fatty acid profiles. J. Chromatogr. B, 2004, 813: 53-58
40. C Wang, H W Kong, Y F Guan et al. Plasma Phospholipid Metabolic Profiling and Biomarkers of Type 2 Diabetes Mellitus Based on High-Performance Liquid Chromatography/Electrospray Mass Spectrometry and Multivariate Statistical Analysis .Anal. Chem., 2005, 77: 4108-4116
41. H R Tang, Y L Wang, J C Lindon et al. UKP: 0323451.5
42. SlimRM, Roberstson D G, AjbassamM, et al. Effict od dexamethasone on the metabonomics profile associated with phosphodiesterase inhibitor-induced vascular lesions in rats. Toxicol ApplRas, 2003, 183(2): 108
43. Watkins S M , Reifsnyder P R, Pan H J. et al. lipid metabolomewide effects of the PPAR gamma agonist rosiglitazone. Lipid res,2002, 43 (11): 1809
44. Zhang Z P, Su Y, Yue N, Zhao Y G. Bichemical mechanism of comparesion studies of clozapine and quetiapine by using metabonomic method in rats. Asian J. Drug Metabolism and Pharmacokinetics, 2004, 4 (4): 281
45. J C Lindon, J K Nicholson, E Holmes et al. Contemporary issues in toxicology the role of metabonomics in toxicology and its evaluation by the COMET project.Toxicol. Appl. Pharmacol, 2003, 187: 137-146
46.J C Lindon, H C Keun, T M D Ebbels et al. The Consortium for Metabonomic Toxicology (COMET): aims, activities and achievements. Pharmacogenomics, 2005,6: 691-699
47. Geisow MJ. Proteomics: one small step for a digital computer, one giant leap for humankind. Not Biotechnol.1998, 16(2): 206
48.Eisen MB，Spellman PT，Brown PO，et a1．Ouster analysis and display of genome-wideexpression patterns．Proc Natl Acad Sci USA，1998，95(25)：14863 14868
49. Holmes E，NiehoUs AW，Lindon JC，et a1．Chemometric models for toxicity classification based on NMR spectra ofbiofluids．ChemResToxicol.2OOO，13(6)：471-478
50.Raamsdonk LM，Teusink B，Broadhurst D，et al．A functional genomics strategy that usesme tabolome data to reveal the phenotype of silent mutations．Nat Biotechnol.2001，19(1)：45 50
51.Bandy JG，Spurgeon DJ，Svendsen C．et al．Earthworm species of the genus Eisenia canbe phenotypically differentiated by metabolic profiling．FEBS Lett.2OO2，521(1-3)：115-120．
52.KS Solanky,N J Bailey,B M Beckwith-Hall.et al.Biofluid 1H NMR-based metabonomictechniques in nutrition research metabolic effects of dietary isoflavones in humans.Nutr.Biochem.,2005,16:236-244.
53.YL Wang,HR Tang,JK Nicholson et al.A metabonomic strategy for the detection of themetabolic effects of chamomile(MatricariarecutitaL.)ingestion.Agric FoodChem.2005,53:191-196.
54.贾伟，蒋健，刘平等.代谢组学在中医药复杂理论体系研究中的应用.中国中药杂志，2006，31（8）：621～624.
55.王米渠,张卫,李珉,等.基因芯片筛选针刺衰老鼠涌泉穴的318个差异表达基因初报.中国中西医结合杂志,2002,22(11):928.
56.王米渠,冯韧,严石林等.基因表达谱芯片发现热药疗寒的相关七类基因初报.中医杂志，2003,44(4):228.
57.王建红,罗来成,于润桥.基于系统工程理论的中医方证研究.辽宁中医杂志.2006;33(3):296
58.王文健,沈自尹,张新民,等.补肾法对老年男性下丘脑-垂体-性腺轴作用的临床和实验研究.中医杂志1986;27(4):32
59.蔡定芳,沈自尹,张玲绢,等.右归饮对大鼠下丘脑-垂体-肾上腺-胸腺轴功能抑制模型的影响.中国免疫学杂志1994;10(7):236
60.蔡定芳,沈自尹,陈晓红,等.乌头碱对大鼠下丘脑促肾上腺皮质激素释放激素含量的影响.中国中西医结合杂志1996;16(9):544
61.钟历勇,沈自尹,蔡定芳,等.补肾健脾活血三类复方对下丘脑-垂体-肾上腺-胸腺轴及CRF基因表达的影响.中国中西医结合杂志1997;17(1):39
62.俞瑾,陈红英,毛秋芝.肾主生殖的实验研究.中西医结合杂志1989;9(9):548
63.俞瑾.肾主生殖与生命网络研究中的启示.中国中西医结合杂志2000;20(6):409
64.沈一丁，费贵强，张宇.代谢组学促进中医药现代化.精细化工.2005，22（6）：444-446
65.许国旺，杨军．代谢组学及其研究进展．色谱，2003，21(4)：316 320．p74-76
1.陈主初.病理生理学第 1版.北京:人民卫生出版社.2006:122-139
2.周爱儒.生物化学第 6版.北京:人民卫生出版社.2006:202-347
3.李天心主编,医学心理学,北京:北京医科大学协和医科大学联合出版社,1998:148-159。
4.BornsteinSR,ChrousosGP.Adrenocorticotropin(ACTH) andN0n-ACTH MediatedRegulationoftheAdrenalCortex:NeuralandImmuneInputs.ClinicEndocrinolMetabolism,1999,84(5):1729-1736.
5.亓晓丽,姚树桥.应激与海马可塑性及其机制的研究进展.中国行为医学科学.2003;11(3):356-358
6.McEwen BS. Stress and hippocampal plasticity.Annu Rev Neurosci.1999;22:105-122。
7.张艳美,杨权,许崇涛,等.慢性应激对大鼠海马锥体细胞形态结构的效应.生物化学与生物物理进展.2002;29(5):719-723
8.McEwenBS.Effectsofadverseexperiencesforbrainstructureandfunction.BiolPsychiatry.2000,48(8):721-731
9.VyasA.Chronicstress inducescontrastingpatternsofdendriticremodelinginhippocampalandamygdaloidneurons.JNeurosci.2002,22(15):6810-6818
10.敖海清,徐志伟,严 灿,等.逍遥散对应激大鼠海马突触体内PKC活性及Ca2+浓度的影响.山东中医杂志.2006,25(2):112-114
11.唐已婷,陈家旭.束缚应激与中医肝的关系.中国医药学报.2002;17(2):82-83
12.唐已婷,陈家旭.慢性束缚应激的中枢介质-促皮质释放激素[J].中国行为医学科学,2001,10(4):394-396
13.唐已婷,陈家旭.慢性束缚应激动物模型的研究进展(综述)[J].北京中医药大学学报,2002,25(1):225-28
14.唐已婷,陈家旭.三种中药复方对慢性束缚应激大鼠下丘脑-垂体-肾上腺轴的调节[J].北京中医药大学学报,2002,25(3):23-26
15.赵歆,陈家旭,杨建新,等.疏肝中药复方对慢性束缚应激大鼠下丘脑 垂体 肾上腺轴的调节[J].实验动物科学与管理,2003,20(2):6-12
16.陈家旭,唐已婷.慢性束缚应激模型大鼠相关脑区前阿黑皮素基因表达的变化及中药复方的影响[J].中国药物与临床,2003,3(3):223-226
17.李伟,陈家旭,杨建新,等.疏肝、健脾、补肾复方对慢性束缚应激大鼠行为学和免疫功能的影响[J].中国实验动物学报,2003,11(1):33-37
18.陈家旭,唐已婷.逍遥散对慢性束缚应激模型大鼠相关脑区 CRF基因表达的影响[J].中国应用生理学杂志,2004,20(1):71-74
19.陈家旭,杨建新,赵歆,等.慢性束缚应激大鼠下丘脑β-内啡肽变化及中药复方对其的影响[J].中国医药学报,2004,19(2):83-85
20.陈家旭,李伟,赵歆,等.三种中药复方对慢性束缚应激大鼠行为及皮层和海马 NT3的影响[J].北京中医药大学学报,2004,27(2):19-23
21.马梦茵,陈家旭.论辨证论治的发展方向[J].中国医药学报,2004,19(6):367-369
22.ChenJX,LiW,ZhaoX,YangJX,XuHY,WangZF,YueGX.ChangesofmRNAexpressionofenkephalinandprodynorphininhippocampusofratswithchronicimmobilizationstress.WorldJGastroenterol.2004;10(17):2547-9
23.岳广欣,陈家旭,王竹风.肝主疏泄的生理学基础探讨[J].北京中医药大学学报,2005,02:1-4.
24.陈家旭,李伟,赵歆,等.慢性束缚应激大鼠海马脑啡肽mRNA和前强啡肽mRNA表达及中药复方的影响[J].中国应用生理学杂志,2005,21(2):121-125
25.王竹风,陈家旭,岳广欣,等.刍议肝郁理论及其影响[J].中华中医药杂志,2005,20(6):367-368
26.岳广欣,陈家旭,王竹风.中医心肝肾三脏在应激反应中的作用 [J].辽宁中医杂志,2005,32(6):528-530
27.李伟,陈家旭.慢性束缚应激大鼠海马 BDNFTrkBNT_3的变化及逍遥散对其影响[J].中医药学刊,2005,23(7):1205-1208
28.王竹风,陈家旭,岳广欣.慢性应激与海马神经元的关系探析[J].辽宁中医学院学报,2005,7(4):337-339
29.王 竹 风 ,陈 家 旭 ,赵 歆 ,等 .肝 主 升 发 在 五 脏 中 的 作 用 探 微 [J].辽 宁 中 医 杂 志 ,2005,32(8):776-777
30.王竹风,陈家旭,赵歆,等.讨肝郁的影响[J].时珍国医国药,2005,16(10):948-949
31.徐洪雁,赵歆,陈家旭,等.慢性束缚应激大鼠海马 CA1区 L-ENK的变化及逍遥散的调节作用.世界科学技术-中医药现代化 2005;7(5):18-22
32.王竹风.慢性束缚应激对脑区 NMDA受体及生长相关蛋白的影响和逍遥散的调节作用.北京中医药大学博士研究生学位论文.2006:3
33.岳广欣. 束缚应激大鼠中枢AMPA受体及相关蛋白变化与肝主疏泄的关系.北京中医药大学博士研究生学位论文.2006:130
34.陈家旭,杨维益.神经-内分泌-免疫网络研究概况及其与中医肝脏关系的探讨.北京中医药大学学报.1995;18(4):7-11
35.严灿,徐志伟,李艳,等.调肝、补肾、健脾方药对慢性心理应激大鼠单胺类神经递质影响的比较研究.中国中西医结合杂志.2002;22(12):925-928
36.徐志伟,严灿,李艳,等.慢性心理应激大鼠血浆及中枢β-EP、ACTH含量的变化及调肝治法方药的影响.中药药理与临床.2002;18(5):37-39
37.王朝勋,郑洪新,王继伟,等.怒伤肝与神经 内分泌 免疫系统失调探析.辽宁中医杂志1997;24(5)∶205-206
38.严灿,邓中炎,王剑.调肝方药对束缚应激大鼠神经内分泌免疫功能的调节作用.中国病理生理杂志,2000,16(6):560-562
39.叶晓平,宋纯清.柴胡皂苷药理研究进展.中草药,2004,35(12):1434-1436
40.唐伟军,卢新华,周大现,等.柴胡镇痛效应的研究.湖南中医杂志,2003,19(4):52-53
41.李芳,李建北,张东明.柴胡的药理研究进展.时珍国医国药,2004,15(2):120-121
42.史青,聂淑琴,黄璐琦.柴胡属植物化学成分及药理研究新进展.中国实验方剂学杂志,2002,8(5):53-56
43.刘琳娜,梅其炳,程建峰.当归挥发油研究的进展.中成药,2002,24(8):621-623
44.夏泉,张平,李绍平,等.当归的药理作用研究进展.时珍国医国药,2004,15(3):164-166
45.张艳,明亮,王瑜,等.白芍总甙的抗惊厥作用.中国药理学通报,1994,10(5):372-374.
46.张 鸿 炼 , 吴 文 蔚 . 白 芍 总 甙 的 药 理 作 用 与 临 床 应 用 . 中 国 医 院 药 学 杂 志 ,1995,15(8):371-372
47.周辉,张安平,陈敏珠,等.白芍总甙对应激大鼠下丘脑-垂体-肾上腺轴和免疫功能的调节.中国药理学通报,1994,10(6):429-432
48.邓 兆 智 , 钟 秋 生 . 白 芍 总 苷 的 药 理 与 临 床 研 究 考 释 . 实 用 中 医 内 科 杂 志 ,2004,18(3):181-182
49.邓兆智,钟秋生,刘文国,等.芍药甙对体外培养的大鼠海马和隔区神经元的影响.解剖学研究,2004,24(4):270-272
50.杨翠平,劳业兴,吴凤薇,等.白术的研究进展.中药材,2002,25(3):206-208.
52.徐丽珊,金晓玲,邵邻相.白术及白术多糖对小鼠学习记忆和抗氧化作用的影响.科技通报,2003,19(6):513-515
52.国家中医药管理局《中华本草》编委会.中华本草.第 1版.上海:上海科学技术出版社,1999
53.李素民,杨秀岭,赵智,等.干姜和生姜药理研究进展.中草药,1999,30(6):471-473.
54.梁呈元,李维林,张涵庆,等.薄荷化学成分及其药理作用研究进展.中国野生植物资源,2003,22(3):9-12
1.孟凡胜,隗学玲,沈烈行 .逍遥散药理研究及临床应用进展.国医论坛,2005,20(3):52-53
2.马增春,高月,谭洪玲,梁乾德,王升启.用分子中药组学技术研究四物汤补血的作用机理.世界科学技术中医药现代化,2005,7(3):24-28
3.刘昌孝.中药药代动力学研究的难点和热点. 药学学报, 2005,40(5):395-401
4.李峰,康廷国.系统论与中药复方作用机制探讨.中医药学刊,2003,21(9):1479-1480
5.王本祥,周秋丽.关于中药活性成分的认识及其研究方法.中国中药杂志,2001,26(1):10-13
6.潘卫松,刘美风,石钺,等.血清药理学、血清化学和中药药代动力学.世界科学技术-中药现代化,2002,4(3):53-55
7.姜廷良,霍海如.重视中药多组分整合作用的研究.世界科学技术-中药现代化.2002,5(2):1-3
8.王升启.中药基因组学与中药蛋白质组学.世界科学技术-中药现代化,2000,2(1):4-8
9.Gennan JB，Bauman DE，Burrrin DG，et al.Metabolomics in the opening decade of the 21stcentury:building ther oads to individualized health．J Nutn，2004，134(10)：2729
10.中药现代化科技发展战略研究课题组.中药现代化重大基础理论研究与重点任务.世界科学技术-中药现代化,2001;3(6):1-6
11.王忠,王阶,王永炎.后基因组时代中医证候组学研究的思考.中国中西医结合杂志,2001;8(21):621-623
12.颜贤忠,赵剑宇,彭双清,廖明阳.代谢组学在后基因组时代的作用.波谱学杂志,2004,21(2):263-271
13.Holmes E,Nicholls AW,Lindon JC,Connor SC,Connelly JC,Haselden JN,Damment SJ,Spraul M,Neidig P,Nicholson JK.Chemometric models for toxicity classification basedon NMR spectra of biofluids.Chem Res Toxicol.2000 Jun;13(6):471-8.
14.Waters NJ,Waterfield C J,Farrant R D,et al.Metabonomic Deconvolution Of EmbeddedToxicity:Application To Thioacetamide Hepato-and Nephrotoxicity.Chem Res Toxicol.2005,18(4):639-654.
15.Lindon JC,Nicholson J K,Holmes E,et al.Metabonomics:Metabolic processes studiedby NMR spectroscopy of biofluids.Concepts Magn Reson.2000,12:289-320.
16.王竹风.慢性束缚应激对脑区 NMDA受体及生长相关蛋白的影响和逍遥散的调节作用.北京中医药大学博士研究生学位论文.2006:3
17.岳广欣. 束缚应激大鼠中枢 AMPA受体及相关蛋白变化与肝主疏泄的关系.北京中医药大学博士研究生学位论文.2006:130
18.石慧琳,程义勇,李树田等.慢性束缚应激对大鼠肠粘膜屏障功能的影响.中国行为医学科学.2003,12(3):251-253
19.吕志平,刘承才. 肝郁 大鼠的脂质过氧化反应及逍遥散的保护作用.山东中医学院学报,1995,19(3):199
20.吕志平,刘承才. 肝郁 大鼠血浆TXA2、PGI2水平与肝微循环变化及逍遥散作用.中国微循环,2000,4(3):160
21.唐已婷,陈家旭.束缚应激与中医肝的关系.中国医药学报.2002;17(2):82-83
22.唐 已 婷 ,陈 家 旭 .慢 性 束 缚 应 激 的 中 枢 介 质 -促 皮 质 释 放 激 素 .中 国 行 为 医 学 科学,2001,10(4):394-396
23.唐 已 婷 ,陈 家 旭 .慢 性 束 缚 应 激 动 物 模 型 的 研 究 进 展 (综 述 ).北 京 中 医 药 大 学 学报,2002,25(1):225-28
24.唐已婷,陈家旭.三种中药复方对慢性束缚应激大鼠下丘脑-垂体-肾上腺轴的调节.北京中医药大学学报,2002,25(3):23-26
25.赵歆,陈家旭,杨建新,李伟,刘晓兰,康纯洁,刘燕.疏肝中药复方对慢性束缚应激大鼠下丘脑垂体 肾上腺轴的调节.实验动物科学与管理,2003,20(2):6-12
25.陈家旭,唐已婷.慢性束缚应激模型大鼠相关脑区前阿黑皮素基因表达的变化及中药复方的影响.中国药物与临床,2003,3(3):223-226
27.李伟,陈家旭,杨建新,赵歆,唐已婷,刘晓兰,徐洪雁.疏肝、健脾、补肾复方对慢性束缚应激大鼠行为学和免疫功能的影响.中国实验动物学报,2003,11(1):33-37
28.陈家旭,唐已婷.逍遥散对慢性束缚应激模型大鼠相关脑区 CRF基因表达的影响.中国应用生理学杂志,2004,20(1):71-74
29.陈家旭,杨建新,赵歆,李伟,陈青红,马梦茵,李柳骥,汪南玥,薛飞飞,洪梅.慢性束缚应激大鼠下丘脑β-内啡肽变化及中药复方对其的影响.中国医药学报,2004,19(2):83-85
30.陈家旭,李伟,赵歆,杨建新,陈青红,马梦茵,李柳骥,徐洪雁.三种中药复方对慢性束缚应激大鼠行为及皮层和海马 NT3的影响.北京中医药大学学报,2004,27(2):19-23
31.马梦茵,陈家旭.论辨证论治的发展方向.中国医药学报,2004,19(6):367-369
32.Chen JX,Li W,ZhaoX ,etall.Changes of mRNA expression of enkephalin and prodynorphin in hippocampus of rats with chronic immobilization stress.World J Gastroenterol.2004;10(17):2547-9
33.岳广欣,陈家旭,王竹风.肝主疏泄的生理学基础探讨.北京中医药大学学报,2005,02:1-4
34.陈家旭,李伟,赵歆,杨建新,徐洪雁.慢性束缚应激大鼠海马脑啡肽mRNA和前强啡肽mRNA表达及中药复方的影响.中国应用生理学杂志,2005,21(2):121-125
35.王竹风,陈家旭,岳广欣等.刍议肝郁理论及其影响.中华中医药杂志.2005,20(6):367-368
36.岳广欣,陈家旭,王竹风.中医心肝肾三脏在应激反应中的作用.辽宁中医杂志.2005,32(6):528-530
37.李伟,陈家旭.慢性束缚应激大鼠海马 BDNFTrkBNT_3的变化及逍遥散对其影响.中医药学刊,2005,23(7):1205-1208
38.王 竹 风 ,陈 家 旭 ,岳 广 欣 .慢 性 应 激 与 海 马 神 经 元 的 关 系 探 析 .辽 宁 中 医 学 院 学报,2005,7(4):337-339
39.王竹风,陈家旭,赵歆,岳广欣,汪宝军.肝主升发在五脏中的作用探微.辽宁中医杂志,2005,32(8):776-777
40.王竹风,陈家旭,赵歆,岳广欣,汪宝军.讨肝郁的影响.时珍国医国药,2005,16(10):948-949
41.徐洪雁,赵歆,陈家旭,马梦茵,王竹风,岳广欣.慢性束缚应激大鼠海马 CA1区 L-ENK的变化及逍遥散的调节作用.世界科学技术-中医药现代化 2005;7(5):18-22
42.张艳美.慢性应激、大脑损害与抑郁症.国外医学-精神病分册.2001,28(2):105-109
43.王良斌,张维宁.大鼠不同脑区突触体钙水平的年龄差异.动物学研究,1995,16(4):373-377
44.耿战辉,程义勇,洪燕等.心理应激时大鼠海马神经元的钙状态变化及锌的作用研究.营养学报,2003,5(1):65-68
45.牛玉杰,石年.溴氰菊酯对大鼠神经细胞内游离钙的影响.卫生毒理学杂志,2001,15(4):216-218
46.William M.Vasopressin-induced protein kinase C-dependent superoxide generation contributes to ATP-seneitive potassium channel butnot calcium-sensitive potassium channelfunction impairment after brain injury.Stroke,2001,32(6):1408-1414
47.敖海清,徐志伟,严灿等.逍遥散对应激大鼠海马突触体内PKC活性及Ca2+浓度的影响，山东中医杂志.2006,25（2）:112-114
48.MacManus J P,Hill IE,Preston E,et al.Difference in DNA fragmentation followingTransient cerebral or decapitation ischemia in rat.J Cereb Blood Flow Metab,1995,15(5):728-737