维吾尔医异常黑胆质性糖尿病病症模型代谢组学研究
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摘要
目的:本研究在建立维医异常黑胆质性糖尿病病症大鼠模型基础上运用基于核磁共振的代谢组学技术,对异常黑胆质证、糖尿病、异常黑胆质性糖尿病病证大鼠及正常大鼠血清和尿液的代谢组分加以分析、判断及发现差异性生物标记物,探讨异常黑胆质性糖尿病病证的物质代谢网络,探索异常黑胆质在糖尿病发生发展过程中的病理生理基础及作用。方法:根据维吾尔医学理论建立维吾尔医异常黑胆质性糖尿病病证模型,对其血清和尿液进行核磁共振氢谱检测,对采集谱图采用OPLS-DA分析进行数据解读。通过皮尔森相关系数显著性差异检测及二维谱技术确定各组大鼠血清和尿液中的差异性代谢成分。结果:不同大鼠血清和尿液1HNMR代谢谱存在差异,血清中有VLDL、LDL、不饱和脂肪酸、丙酮、乙酰乙酸、丙酮酸、柠檬酸、甘氨酸、丙氨酸、酪氨酸、苯丙氨酸、亮氨酸、异亮氨酸、瓜氨酸、蛋氨酸、肌酸、肌醇、鲨肌醇、尿素等差异性代谢物出现(P<0.05),不同大鼠尿液差异性代谢物有柠檬酸、胍基乙酸、酪氨酸、蛋氨酸、赖氨酸、苯丙氨酸、色氨酸、异亮氨酸、缬氨酸、丙氨酸、天冬氨酸、肌酸酐等(P<0.05)。结论:异常黑胆质证组、糖尿病组、异常黑胆质性糖尿病病证组、正常对照组大鼠血清和尿液间存在差异代谢组分,异常黑胆质证的发生、发展过程中脂代谢紊乱,糖尿病大鼠存在糖代谢及能量代谢障碍。糖代谢紊乱、肌酸合成障碍等是糖尿病大鼠与异常黑胆质性糖尿病病症大鼠共同的病理生理过程,不同组大鼠间代谢组分差异在尿液中更明显。
Objective: based on the establishment of Abnormal savda disease rat model with Diabetes Mellitus,appling the metabonomics technology through nuclear magnetic resonance (NMR),, analyse and discover the biological marker by Metabonomics study of Serum and urine of Abnormal savda model group,diabetes Mellitus model group, Abnormal savda disease carring Diabetes Mellitus model group and control group,investigate the substance metabolism network of Abnormal savda disease earring Diabetes Mellitus, explore the Pathophysiological background and function in the process of Abnormal savda disease carring Diabetes Mellitus.
     Methods: According to the theory of Uighur traditional medicine, Abnormal savda disease carring Diabetes Mellitus rat model was established, A 1H NMR based metabonomic analysis was performed on these serum and urine samples. A multi-steps date preprocessing procedure was completed and then Orthogonal Partial Least-Squares Discriminant Analysis(OPLS-DA) was fulfilled to distinguish the metabolic phenotypes, the influential biomarkers accounting for the metabolic signature perturbation were discovered by Pearson's product moment correlation coefficient and 2D NMR.
     Results: It was found that between the H NMserum 5'-nucleotidase of four group exist signification difference, the variability of metabolin in the serum of four group were VLDL, LDL, unsaturated fatty acid, acetone, acetoacetic acid, pyruvic acid, citric acid, glycine, alanine, tyrosine, Phe acid, leucine, isoleucine, citrulline, methionine, creatine, inositol, shark inositol, urea,etc. (P<0.05), the variability of metabolin in the urine of four group were citric acid, guanidine acetic acid, tyrosine, methionine, lysine, phenylalanine, tryptophan, isoleucine, valine, alanine, aspartic acid, creatinine, etc. (P<0.05).
     Conclusion: the metablolite in the serum and urine of those Abnormal savda, diabetes, Abnormal savda disease carring Diabetes Mellitus and control group were significant differences.the development of abnormal savda can cause the metabolism disorganization of lipid.the glycolysis and energy synthesis metabolisim disorder in the Diabetes group.glycometabolic disturbance, creatine dyssynthesis are the congenerous Pathophysiological process of Abnormal savda and diabetes group.the sianificant difference of the variability of metabolin in the urine is more than the sianificant difference of the variability of metabolin in the serum.
引文
[1]买买提依明·沙比尔等,维吾尔医诊断学(维吾尔文)[M]乌鲁木齐:新疆科技卫生出版社,1985:17-33
    [2]穆巴拉克、派祖拉、亚尔买买提,等.维吾尔医学中关于体液的探索[J].中国民族民间医药杂志.2002,(55):69-70.
    [3]阿吉·阿布里米提,玉素甫,主编.维吾尔医学基础理论[M].乌鲁木齐:新疆人民卫生出版社,1988:23-134.
    [4]新疆维吾尔自治区卫生厅中国医学百科全书维吾尔医分卷办公室.中国医学百科全书·维吾尔医学分卷[M].新疆:新疆人民卫生出版社,1985:17-25.
    [5]哈木拉提·吾甫尔.维医气质、体液轮及其现代研究[M].乌鲁木齐:新疆科技卫生出版社,2003.
    [6]库热西江·托乎提,阿不都热依木·玉素甫,哈木拉提·吾甫尔.维吾尔医正常体液与异常体液分型比较研究[J].中国民族医药杂志,2004(3):3-4.
    [7]哈木拉提·吾甫尔,阿不都热依木·玉苏甫,吐尔逊·吾甫尔.维医异常体液分型与年龄、性别关系的探讨[J].中国民族民间医药杂志,2003,61:84-86.
    [8]Tang H R,Wang Y L. Metabonomics: a revolution in progress. Prog Biochem Biophys,2006,33(5):401-417
    [9]Nicholls A W, Holmes E, Lindon J C, et al. Metahonomic investigations into hydrazine toxicity in the rat[J]. Chem Res Toxicol,2001,14(8):975-987.
    [10]Steinhauser D, Kopka J.Methods, applicationsand concepts of metabolite profiling: primarymetabolism. EXS 2007,97:171-194
    [11]Robertson DG, Reily MD, Baker JD. Metabonomicsin pharmaceutical discovery and development. JProteome Res 2007,6:526-539
    [12]LenzEM,WilsonID.Analyticalstrategiesinmetabonomics.JProteome Res 2007,6:443-458
    [13]Larive CK. Metabonomics, metabolomics, andmetabolic profiling. Anal Bioanal Chem 2007,387:523
    [14]Dieterle F, Ross A, Schlotterbeck G, Senn H.Probabilistic quotient normalization as robustmethod to account for dilution of complexbiological mixtures. Application in 1H NMRmetabonomics. Anal Chem 2006,78:4281-4290
    [15]Lindon JC, Holmes E, Nicholson JK. Metabonomicstechniques and applications to pharmaceuticalresearch & development. Pharm Res 2006; 23:1075-1088
    [16]Nicholsn J K, Lin don J C, Holmes E.'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica 1999; 29:1181-1189
    [17]Bermensen MF. Erikson NM. Mortens∞LS, et sl. Cau the SlyⅧic index(G1)be used ∞-tool in the prevention sad management of type 2 d "sbntes?[J]. RevⅨab 吼 Stud。 2008,3(2):61-71.
    [18]Nicholson J K, Oflynn M P, Sadler P J. Proton nuclear magnetic resonance studies of serum, plasma and urine f rom fasting normal and diabetic subject s [J].Biochem J,1984,217:365-375.
    [19]Wang C, Kong H W, Guan Y F, et al. Plasma phospholipid metabolic profiling and biomarkers of type 2 diabe2 tes mellitus based on high performance liquid chromatography/elect rospray mass spect rometry and multivariate statistical analysis [J].Anal Chem,2005,77(13):4108-4116.
    [20]Yuan K L, Kong H W, Guan Y F,et al.A GC2 based metabonomics investigation of type 2 diabetes by organic acids metabolic profile [J]. J Chromatogr B,2007,850(1-2):236-240.
    [21]M akinen V2P, Soininen P, Forsblom C,et al.Diagnosing diabetic nephropat hy by 1 H NMR metabonomics of serum [J]. Mag Res Mat Phy,2006,19:281-296.
    [22]Jin E S,Burgess S C,Merritt M,et al.Differing mechanisms of hepatic glucose overproduction in t riiodot hy2 roninet reated rats vs. Zucker diabetic fatty rats by NMR analysis of plasma glucose [J].Am J Physiol Endocri2 nol Metab,2005,288:E654-E662.
    [23]董继扬,徐乐等.代谢组学数据分析方法及在糖尿病研究中的应用[J].波谱学杂志,2007,12(4):381-393。
    [24]Beaufour C C, Ballon N, Le Bihan C, et al. Effects of antidepressants in operant conflict procedure of anxiety in the rat [J]. Pharmacol Biochem Behavior,1999,62(4): 591-599.
    [25]张莉,哈木拉提·吾甫尔等.异常黑胆质载体动物肿瘤模型的建立及其神经内分泌免疫指标的改变[J].国际病理科学与临床杂志.2007,27(5):376-381
    [26]哈木拉提·吾甫尔,阿依努尔·买提斯迪克,努尔买买提·艾买提,等.异常黑胆质证载体动物模型的建立及其自然恢复反证[J].新疆医科大学学报,2006,29(10):910-914.
    [27]张有辉1,玉苏甫·吐尔逊2,哈木拉提·吾甫尔,等.异常黑胆质性肝癌病证模型肝硬化期免疫、内分泌紊乱状态[J].科技导报,2010,28(11):39-42
    [28]施红,雅,余文珍,等.石斛合剂改善模型大鼠胰岛素抵抗的实验研究[J].福建中医学 院学报,2004,14(6):24-27
    [29]郭啸华,刘志红,李恒,等.高糖高脂饮食诱导的2型糖尿病大鼠模型及其肾病特点[J].中国糖尿病杂志,2002,]0(5):290-294
    [30]赵宝珍,白秀平,荣青峰,等.实验2型糖尿病大鼠模型的研究[J].中国药物与临床,2002,2(6):383-385
    [31]洪丽莉,许冠荪,申国明,等.SD大鼠2型糖尿病模型的建立[J].实验动物科学与管理,2005,22(4):1-3
    [32]刘欣秋,雷鸣.实验性非胰岛素依赖性糖尿病模型探讨[J].黑龙江医药科学,2001,24(3)
    [33]巴吐·尔·买买提明,阿仙姑·哈斯木,陈春丽,等.基于不同模式识别方法的大鼠尿液核磁共振氢谱分析[J]新疆医科大学学报,2010,33(7):755-761
    [34]董继扬,徐乐,曹红婷,等.代谢组学数据分析方法及在糖尿病研究中的应用[J].波谱学杂志.2007,24(4):181-193
    [35]严兴海,巴吐尔·买买提明,李春燕等.基于核磁共振的肾虚痰瘀型哮喘患者血浆的代谢组学研究[J]新疆医科大学学报.2010,33(3):228-233
    [36]易沙克江·马合穆德.中国医学百科全书·维吾尔医学[M].上海:上海科学技术出版社,2005
    [37]热娜古丽·艾则孜,沙吉旦·阿不都热衣,金伟,等.复杂性疾病维吾尔医分型及其血栓前状态研究[J].中国中医药信息杂志,2009,2(16)23-25
    [38]Tang H R,Wang Y L. Metabonomics :a revolution in progress. Prog Biochem Biophys, 2006,33(5):401-417
    [39]VIANTM, ROSENBLUM E,TJEERDEMA R.[J]. Environmental Science & Technology,2003,37(21):4982-4989.
    [40]KR ISHNAN P, KRUGER N J, RATCL IFFE R G. [J].Journal of Experimental Botany,2005,56(410):255-265.
    [41]LE GALL G, COLQUHOUN I J, DAV IS A L, et al[J].Journal of Agriculture and Food Chemistry,2003,51(9):2447-2456.
    [42]PAN Z Z, RAFTERY D[J]. Analytical and Bioanalytical Chemistry,2007, 387(2):525-527.
    [43]RATCL IFFE R G, SHACHAR2H ILL Y[J]. Annual Review of Plant Physiology and PlantMolecularBiology,2001,52(1):499-526.
    [44]VERPOORTE R, CHO I Y H, KIM H K. [J]. Phytochemistry Reviews,2007, 6(1):3-14.
    [45]GR IFF IN J[J]. CurrentOp inion in Chemical Biology,2003,7(5):648-654.
    [46]周爱儒,查锡良.生物化学[M].北京:人民生出版社,2004.
    [47]艾斯克尔·吐拉洪,哈木拉提·吾甫尔,豪富华等.基于NMR的维开尔医异常黑胆质型肿瘤患者血浆代谢组学研究[J].科技导报,2009,27(13):27-31
    [48]刘晓永,钱和,刘建利.肌醇研究近况与展望[J].江苏食品与酵,2003,30(4):7-10.
    [49]Yu J, Berggren PO, Barker CJ. An autocrine insulin2feedback loop maintains pancreatic β2cell 2Phosphorylated inositol lipids[J]. Mol Endocrinol,2007,21(11):277522784
    [50]Ostlund RE J r, McGill JB, Herskowitz I et al. D2chiro2inositol metabolism in diabetes mellitus[J].rocnatl Acad Sci,1993,90:9988-9992.
    [51]舒向荣,王霆,程泽能,等.红杉醇等肌醇类物质降血糖作用机制的研究[J].中南药学2009,1(7):43-46
    [1]Steinhauser D, Kopka J. Methods, applicationsand concepts of metabolite profiling: primarymetabolism. EXS 2007; 97:171-194
    [2]Robertson DG, Reily MD, Baker JD. Metabonomicsin pharmaceutical discovery and development. JProteome Res 2007; 6:526-539
    [3]LenzEM,WilsonID.Analyticalstrategiesinmetabonomics.JProteome Res 2007; 6: 443-458
    [4]Larive CK. Metabonomics, metabolomics, andmetabolic profiling. Anal Bioanal Chem 2007; 387:523
    [5]Dieterle F, Ross A, Schlotterbeck G, Senn H.Probabilistic quotient normalization as robustmethod to account for dilution of complexbiological mixtures. Application in 1H NMRmetabonomics. Anal Chem 2006; 78:4281-4290
    [6]Lindon JC, Holmes E, Nicholson JK. Metabonomicstechniques and applications to pharmaceuticalresearch & development. Pharm Res 2006; 23:1075-1088
    [7]Nicholsn J K, Lin don J C, Holmes E.'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica 1999; 29:1181-1189
    [8]Fiehn O. Metabolomics--the link between genotypes and phenotypes. Plant Mol Biol 2002;48:155-171
    [9]Tang H R,Wang Y L. Metabonomics: a revolution in progress. Prog Biochem Biophys,2006,33(5):401-417
    [10]Holmes E, Tang H R, Wang Y L, et al, The assessment of plant metabolite profiles by NMR-based methodologies.Plant Med,2006,72(9):771-785
    [11]Taylor J,King R D,Altmann T,Fiehn O.Bioinformatics,2002,18:241
    [12]VIANTM, ROSENBLUM E, TJEERDEMA R. [J]. Environmental Science & Technology,2003,37 (21):4982-4989.
    [13]KR ISHNAN P, KRUGER N J, RATCL IFFE R G. [J]. Journal of Experimental Botany,2005,56 (410):255-265.
    [14]LE GALL G, COLQUHOUN I J, DAV IS A L, et al. [J]. Journal of Agriculture and Food Chemistry,2003,51(9):
    [15]2447-2456.
    [16]PAN Z Z, RAFTERY D. [J]. Analytical and Bioanalytical Chemistry,2007,387 (2): 525-527.
    [17]RATCL IFFE R G, SHACHAR2H ILL Y. [J]. Annual Review of Plant Physiology and PlantMolecularBiology,2001,52 (1):499-526.
    [18]VERPOORTE R, CHO I Y H, KIM H K. [J]. Phytochemistry Reviews,2007,6(1): 3-14.
    [19]GR IFF IN J. [J]. CurrentOp inion in Chemical Biology,2003,7 (5):648-654.
    [20]Lauridsen M, Hansen SH, Jaroszewski JW, Cornett C. Human urine as test material in 1H NMR-based metabonomics: recommendations for sample preparation and storage. Anal Chem 2007; 79:1181-1186
    [21]Ott KH, Aranibar N. Nuclear magnetic resonance metabonomics: methods for drug discovery and development. Methods Mol Biol 2007; 358:247-271
    [22]Serkova NJ, Niemann CU. Pattern recognition and biomarker validation using quantitative lH-NMRbased metabolomics. Expert Rev Mol Diagn 2006; 6:717-731
    [23]Constant inou MA, Tsant i 1 i-Kakoul idou A,Andreadou I, Iliodromitis EK, Kremastinos DT, Mikros E. Appl i cat ion of NMR-bas e d metabonomics in the investigation of myocardial ischemia-reperfusion, ischemic preconditioning and antioxidant intervention in rabbits. Eur J Pharm Sci 2007; 30:303-314
    [24]Lindon JC, Holmes E, Nicholson JK. Metabonomics techniques and applications to pharmaceutical research & development. Pharm Res,2006,23(6):1075-1088.
    [25]Cheng LL, Chang IW, Louis DN, Gonzalez RG. Correlation of high-resolution magic angle spinning proton magnetic resonance spectroscopy with histopathology of intact human brain tumour specimens. Cancer Res,1998,58(9):1825-1832.
    [26]Price KE, Vandaveer SS, Lunte CE, Larive CK. Tissue targeted metabonomics: metabolic profiling by microdialysis sampling and microcoil NMR. J Pharm Biomed Anal,2005,38(5):904-909.
    [27]Bollard ME, Garrod S, Holmes E, Lindon JC, Humpfer E, Spraul M, Nicholson JK. High-resolution 1H and 1H-13C magic angle spinning NMR spectroscopy of rat liver. Magn Reson Med,2000,44(2):201-207. [DOI]
    [28]LENZ EM, W ILSON ID. [J]. Journal of Proteome Research,2007,6 (2):443-458.
    [29]HALKET JM, WATERMAN D, PRZYBOROWSKA AM, et al. [J]. Journal of ExperimentalBotany,2005,56(410):219-243.
    [30]ROESSNER U, WAGNER C, KOPKA J, et al. [J]. The Plant Journal,2000,23 (1): 131-142.
    [31]MAURER H H. [J]. J Chromatogr: B: Biomedical Sciences and App lications,1998, 713 (1):3-25.
    [32]BENDAHL L, HANSEN S H, GAMMELGAARD B, et al. [J]. Journal of Pharmaceutical and Biomedical Analysis,2006,40 (3):648-652.
    [33]CASTRO2PEREZ J, PLUMB R, GRANGER J H, et al. [J]. Rap id Communications inMass Spectrometry,2005,19(6):843-848.
    [34]JOHNSON K A, PLUMB R. [J]. Journal of Pharmaceutical and BiomedicalAnalysis, 2005,39 (3/4):805-810.
    [35]LEANDRO C C, HANCOCK P, FUSSELL R J, et al. [J]. J Chromatogr: A,2006, 1103(1):94-101.
    [36]TOLSTIKOV V, F IEHN O. [J]. Analytical Biochemistry,2002,301 (2):298-307.
    [37]PERRETT D, ALFRZEMA L, HOWSM, et al. [J]. Biochemical Society Transactions,1997,25 (11):273-278.
    [38]PERRETT D, ROSS G. [J]. Trac2Trends in Analytical Chemistry,1992,11 (4):156-163.
    [39]TERABE S, MARKUSZEWSKIM, INOUE N, et al. [J]. Pure and App lied Chemistry,2001,73 (10):1563-1572.
    [40]BR ITZ2MCKIBB IN P, TERABE S. [J]. Chemical Record,2002,2 (6):397-404.
    [41]SOGA T, IMA IZUM IM. [J]. Electrophoresis,2001,22 (16):3418-3425.
    [42]SOGA T, OHASH I Y, UENO Y, et al. [J]. Journal of Proteome Research,2003,2 (5):488-494.
    [43]SOGA T, UENO Y, NARAOKA H, et al. [J]. Anal Chem,2002,74 (10):2233-2239.
    [44]KELL D B. [J]. CurrentOp inion inMicrobiology,2004,7 (3):296-307.
    [45]D IEM M, BOYDSTON-WH ITE S, CH IR IBOGA L. [J]. App lied Spectroscopy, 1999,53(4):148A-161A.
    [46]Yang J,GW,Kong H W,Zheng Y F,Pang T,Yang Q.Jchromatogr B,2002,780:27
    [47]Beaudry F, Le Blanc J C Y, Coutu M,Ramier I,Moreu J P, Brown N K.Biomed Chromatogr,1999,13:363
    [48]Oliver D J, Nikolau B, Wurtele E S.Metabolic Engineering,2002,4:98
    [49]BRAZMA A, H INGAMP P, QUACKENBUSH J. [J]. Nature Genetics,2001,29 (4):365-371.
    [50]Nobeli I, Thornton JM. A bioinformatician's view of the metabolome. Bioessays, 2006,28(5):534-545. [DOI]
    [51]Robertson DG. Metabonomics in toxicology: a review. Toxicol Sci,2005,85(2): 809-822.
    [52]Trygg J, Holmes E, Lundstedt T. Chemometrics in metabonomics. J Proteome Res, 2007,6(2):469-479. [DOI]
    [53]Raamsdonk L M, Teusink B, Broadhurst D, et al. A functionalgenomics strategy that uses metabolome data to revealthe phenotype of silent mutations. Nat Biotechnol,2001,19(1):45-50
    [54]Nicholson J K, Connelly J, Lindon J C, et al. Metabonomics:a platform for studying drug toxicity and gene function. Nat Rev Drug Discov,2002,1(2):153-161
    [55]Lindon JC, Nicholson JK, Everett JR. NMR spectroscopy of biofluids. Ann Rep NMR Spect,1999,38:1-88.
    [56]Ellis DI, Dunn WB, Griffin JL, Allwood JW, Goodacre R. Metabolic fingerprinting as a diagnostic tool. Pharmacogenomics,2007,8(9):1243-1266.
    [57]Van Ravenzwaay B, Cunha GC, Leibold E, Looser R, Mellert W, Prokoudine A, Walk T, Wiemer J. The use of metabolomics for the discovery of new biomarkers of effect. Toxicol Lett,2007,172(1-2):21-28.

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