枸杞多糖作用于2型糖尿病大鼠的血清代谢组学研究
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摘要
建立适用于血清代谢物谱研究的气相色谱-飞行时间质谱技术,并用于对照组(NC组)、2型糖尿病模型组(type 2 diabetic model group,DM组)及枸杞多糖干预DM大鼠组(Lycium barbarum polysaccharides,LBP组)大鼠血清代谢物谱的分析。采用主成分分析、正交偏最小二乘判别法等模式识别方法对NC组、DM组及LBP组大鼠血清代谢物谱进行分类,并从血清中寻找与2型糖尿病相关结果部分并未出现显著性分析的潜在生物标志物。结果表明,所建立的方法能将3组大鼠血清代谢物谱分离,DM组大鼠血清中丙氨酸、胸腺嘧啶脱氧核苷酸相对于NC组大鼠降低;LBP干预DM大鼠1个月后,LBP组大鼠血清中丙氨酸、胸腺嘧啶核苷酸含量有所上升。血清中这丙氨酸、胸腺嘧啶核苷酸可能与氨基酸代谢和核苷酸代谢有关,LBP组与DM组大鼠血清中丙氨酸和胸腺嘧啶核苷酸水平的变化体现了大鼠体内氨基酸和核苷酸代谢的改变。本实验可为进一步研究LBP的降糖作用机制提供参考。
Gas chromatography-time of flight-mass spectrometry(GC-TOF-MS) was employed for serum metabolite profiling of rats in the control group(NC), type 2 diabetic model group(DM) and Lycium barbarum polysaccharides(LBP group) treatment groups. Principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used for clustering serum metabolite profiles and finding potential biomarkers. The results demonstrated that the three groups were successfully separated and serum alanine and thymidine were decreased in the DM group as compared to the NC group. One-month intervention with LBP could increase the levels of alanine and thymidine in serum of type 2 diabetic rats, which may be associated with their amino acid and nucleotide metabolism. These results can provide a reference for further study of the hypoglycemic mechanism of LBP.
Gas chromatography-time of flight-mass spectrometry(GC-TOF-MS) was employed for serum metabolite profiling of rats in the control group(NC), type 2 diabetic model group(DM) and Lycium barbarum polysaccharides(LBP group) treatment groups. Principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) were used for clustering serum metabolite profiles and finding potential biomarkers. The results demonstrated that the three groups were successfully separated and serum alanine and thymidine were decreased in the DM group as compared to the NC group. One-month intervention with LBP could increase the levels of alanine and thymidine in serum of type 2 diabetic rats, which may be associated with their amino acid and nucleotide metabolism. These results can provide a reference for further study of the hypoglycemic mechanism of LBP.
引文
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[27]皮子凤,门丽慧,张静,等.五味子治疗大鼠糖尿病肾病作用机制的血清代谢组学研究[J].分析化学,2015,43(2):169-175.DOI:10.11895/j.issn.0253-3820.140564.
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[29]宁萌,李田乐,段大为,等.2型糖尿病早期大鼠血清代谢组学分析[J].生物医学工程与临床,2016,20(2):131-135.
[30]NEWGARD C B,AN J,BAIN J R,et al.A branched-chain amino acid-related metabolic signature that differentiates obese and lean humans and contributes to insulin resistance[J].Cell Metabolism,2009,9(4):311-326.DOI:10.1016/j.cmet.2009.02.002.
[2]贾伟.医学代谢组学[M].上海:上海科学技术出版社,2011:1-16.
[3]JIYE A,TRYGG J,GULLBERG J,et al.Extraction and GC/MS analysis of the human blood plasma metabolome[J].Analytical Chemistry,2005,77(24):8086-8094.DOI:10.1021/ac051211v.
[4]XIAO Y P,WU T X,HONG Q H,et al.Response to weaning and dietary l-glutamine supplementation:metabolomic analysis in piglets by gas chromatography/mass spectrometry[J].Journal of Zhejiang University:Science B,2012,13(7):567-578.DOI:10.1631/jzus.B1100343.
[5]BIRKEMEYER C,KOLASA A,KOPKA J.Comprehensive chemical derivatization for gas chromatography-mass spectrometry-based multi-targeted profiling of the major phytohormones[J].Journal of Chromatography A,2003,993(1/2):89-102.DOI:10.1016/S0021-9673(03)00356-X.
[6]SCHMELZ E A,ENGELBERTH J,ALBORN H T,et al.Simultaneous analysis of phytohormones,phytotoxins,and volatile organic compounds in plants[J].Proceedings of the National Academy of Sciences,2003,100(18):10552-10557.DOI:10.1073/pnas.1633615100.
[7]CHI A P,ZHANG Y,KANG Y J,et al.Metabolic mechanism of a polysaccharide from Schisandra chinensis to relieve chronic fatigue syndrome[J].Journal of Biological Macromolecules,2016,93:322-332.DOI:10.1016/j.ijbiomac.2016.08.042.
[8]NI Y,SU M M,QU Y P,et al.ADAP-GC 3.0:improved peak detection and deconvolution of co-eluting metabolites from GC/TOF-MS data for metabolomics studies[J].Analytical Chemistry,2016,88(17):8802-8811.DOI:10.1021/acs.analchem.6b02222.
[9]WANG G M,SHI D D,PENG Z X,et al.Study on mechanism of costunolide-induced apoptosis in breast cancer MCF-7 cells[J].Chinese Journal of Analytical Chemistry,2015,43(5):682-688.
[10]郭琦.枸杞多糖的提取、分离纯化、溶液性质及其结构的初步研究[D].西安:陕西师范大学,2012:1-10.
[11]孙桂菊,张勇,黄杰,等.枸杞多糖对Ⅱ型糖尿病大鼠肾脏保护作用及其机制研究[J].营养学报,2006,28(1):47-50.DOI:10.3321/j.issn:0512-7955.2006.01.013.
[12]ZOU S,ZHANG X,YAO W B,et al.Structure characterization and hypoglycemic activity of a polysaccharide isolated from the fruit of Lycium barbarum L.[J].Carbohydrate Polymers,2010,80:1161-1167.DOI:10.1016/j.carbpol.2010.01.038.
[13]孙桂菊,王少康,张勇,等.枸杞多糖、茶叶多糖混合物对Ⅱ型糖尿病大鼠降血糖作用及对糖尿病并发症相关指标的影[J].食品研究与开发,2003,24(2):75-78.DOI:10.3969/j.issn.1005-6521.2003.02.033.
[14]LUO Q,CAI Y,YAN J,et al.Hypoglycemic and hypolipidemic effects and antioxidant activity of fruit extracts from Lycium barbarum[J].Life Science,2004,76(2):137-149.DOI:10.1016/j.lfs.2004.04.056.
[15]LI X M.Protective effect of Lycium barbarum polysaccharides on streptozotocin-induced oxidative stress in rats[J].International Journal of Biological Macromolecules,2007,40:461-465.DOI:10.1016/j.ijbiomac.2006.11.002.
[16]蔡慧珍,刘福康,卢慧霞,等.枸杞多糖对2型糖尿病病人的短期干预[J].食品科学,2012,33(13):259-262.
[17]田晓华,丛建波,孙存普,等.氧化苦参碱清除·OH自由基作用及对胸腺嘧啶核苷酸辐射防护作用的ESR研究[J].解放军预防医学杂志,1996,14(6):412-415.
[18]陈忱,蔡慧珍,唐华丽,等.枸杞多糖的组成分析及其荧光标记研究[J].时珍国医国药,2014,25(10):2312-2315.DOI:10.3969/j.issn.1008-0805.
[19]刘源才,孙细珍,许银,等.枸杞多糖组成及含量测定方法的改进[J].食品科学,2013,34(12):292-295.DOI:10.7506/spkx1002-6630-201312060.
[20]蔡慧珍.枸杞多糖对人胰岛素抵抗及血脂的干预作用及其机制[D].南京:东南大学,2012:38-42.
[21]宗灿华,田丽梅.枸杞多糖对2型糖尿病大鼠胰岛素抵抗及脂联素基因表达的影响[J].中国康复理论与实践,2008,14(6):531-532.DOI:10.3969/j.issn.1006-9771.2008.06.014.
[22]TANG H L,CHEN C,WANG S K,et al.Biochemical analysis and hypoglycemic activity of a polysaccharide isolated from the fruit of Lycium barbarum L.[J].International Journal of Biological Macromolecules,2015,77:235-242.DOI:10.1016/j.ijbiomac.2015.03.026.
[23]JIA W P.Diabetes:a challenge for China in the 21st Century[J].Lancet Diabetes Endocrinol,2014,2(4):6-7.DOI:10.1016/S2213-8587(14)70027-0.
[24]ZIMMET P Z,MAGLIANO D J,HERMAN W H,et al.Diabetes:a 21st Century challenge[J].Lancet Diabetes Endocrinol,2014,2(1):56-64.DOI:10.1016/S2213-8587(13)70112-8.
[25]BAO Yuqian,ZHAO Tie,WANG Xiaoyan,et al.Metabonomic variations in the drug-treated type 2 diabetes mellitus patients and healthy volunteers[J].Journal of Proteome Research,2009,8(4):1623-1630.DOI:10.1021/pr800643w.
[26]郭准霄,张俊清,董爱梅.2型糖尿病患者血浆氨基酸浓度的变化及其与血糖和胰岛素的关系[C]//北京医学会糖尿病暨内分泌学会学术年会.北京:北京大学第一医院内分泌科,2012:292-293.
[27]皮子凤,门丽慧,张静,等.五味子治疗大鼠糖尿病肾病作用机制的血清代谢组学研究[J].分析化学,2015,43(2):169-175.DOI:10.11895/j.issn.0253-3820.140564.
[28]张启云,汤喜兰,李冰涛,等.黄连总生物碱对糖尿病大鼠的血浆代谢组学研究[J].中药药理与临床,2013,29(6):42-44.
[29]宁萌,李田乐,段大为,等.2型糖尿病早期大鼠血清代谢组学分析[J].生物医学工程与临床,2016,20(2):131-135.
[30]NEWGARD C B,AN J,BAIN J R,et al.A branched-chain amino acid-related metabolic signature that differentiates obese and lean humans and contributes to insulin resistance[J].Cell Metabolism,2009,9(4):311-326.DOI:10.1016/j.cmet.2009.02.002.