基于超高效液相色谱-四极杆飞行时间质谱联用技术的血瘀模型大鼠血浆代谢组学分析
|
摘要
采用盐酸肾上腺素加冰水浴建立急性血瘀大鼠模型,使用超高效液相色谱-四极杆飞行时间质谱(UPLC-QTOF/MS)检测空白对照组与血瘀模型组中血浆代谢物,用主成分分析(PCA)、有监督偏最小二乘法判别分析(PLS-DA)及正交偏最小二乘法判别分析(OPLS-DA)对代谢组学数据进行多维统计分析,筛选潜在生物标志物。与对照组相比,在血瘀模型组大鼠血浆中检测出46个差异代谢物,血瘀模型组中乙酰胆碱、N6,N6,N6-三甲基-L-赖氨酸、胞嘧啶、乙酰肉碱等21个代谢物显著上调,吲哚丙酸、Lyso PC(14∶0)等25个代谢物显著下调,可能与脂质代谢、半乳糖代谢、亚油酸代谢、不饱和脂肪酸生物合成、糖酵解、花生四烯酸代谢等通路有关。代谢产物可作为血瘀证研究中的重要标记物,该研究结果有助于揭示血瘀证的发病机制,可为临床血瘀疾病的诊断及选用药物治疗提供思路,为后续治疗手段提供参考依据。
Acute blood stasis syndrome was induced in rats by adrenaline hydrochloride and ice water. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry( UPLC-Q-TOF/MS) was conducted on plasma metabolites of normal and model rats. Principal component analysis( PCA),differentiation analysis of supervised partial least squares method( PLS-DA),and orthogonal to partial least squares discriminant analysis( OPLS-DA) on metabolomics data for multidimensional statistical analysis were employed,and the resultingbiomarkers were screened. Compared to the normal group,there were 46 endogenous metabolites in blood stasis-rat plasma. Of these,21 metabolites were significantly upregulated,such as acetylcholine,N6,N6,N6-trimethyl-L-lysine,cytosine,and acetylcarnitine,while 25 metabolites were reduced,including indoleacrylic acid,and lysoPC( 14 ∶ 0). These metabolites were mainly related to metabolic pathways,including lipid metabolism,galactose metabolism,linoleic acid metabolism,biosynthesis of unsaturated fatty acids,glycolysis,and arachidonic acid metabolism. In conclusion,these results indicated that metabolites could be used as important biomarkers for blood stasis syndrome,and could help in revealing the mechanism of blood stasis disease and provide a reference network to determine the disease development stage and appropriate follow-up treatment. Studying altered metabolites in blood stasis model rats can provide insights useful for the diagnosis of blood stasis in the clinic and for the development of drug therapies.
Acute blood stasis syndrome was induced in rats by adrenaline hydrochloride and ice water. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry( UPLC-Q-TOF/MS) was conducted on plasma metabolites of normal and model rats. Principal component analysis( PCA),differentiation analysis of supervised partial least squares method( PLS-DA),and orthogonal to partial least squares discriminant analysis( OPLS-DA) on metabolomics data for multidimensional statistical analysis were employed,and the resultingbiomarkers were screened. Compared to the normal group,there were 46 endogenous metabolites in blood stasis-rat plasma. Of these,21 metabolites were significantly upregulated,such as acetylcholine,N6,N6,N6-trimethyl-L-lysine,cytosine,and acetylcarnitine,while 25 metabolites were reduced,including indoleacrylic acid,and lysoPC( 14 ∶ 0). These metabolites were mainly related to metabolic pathways,including lipid metabolism,galactose metabolism,linoleic acid metabolism,biosynthesis of unsaturated fatty acids,glycolysis,and arachidonic acid metabolism. In conclusion,these results indicated that metabolites could be used as important biomarkers for blood stasis syndrome,and could help in revealing the mechanism of blood stasis disease and provide a reference network to determine the disease development stage and appropriate follow-up treatment. Studying altered metabolites in blood stasis model rats can provide insights useful for the diagnosis of blood stasis in the clinic and for the development of drug therapies.
引文
[1] Li X P. Journal of Changzhi Medical College,2006,20(1):67李晓萍.长治医学院学报,2006,20(1):67
[2] Li W X, Huang M Y, Tang Y P, et al. Chinese Pharm aco lo gical Bulletin,2011,27(12):1761李伟霞,黄美艳,唐于平,等.中国药理学通报,2011,27(12):1761
[3] Wang C,Jia X Y,Hou L W,et al. Acta Pharmaceutica Sinica,2018,53(6):980王翠,贾雪洋,侯璐文,等.药学学报,2018,53(6):980
[4] Li N,Fan X M,Wang Y M,et al. Central South Pharmacy,2014,12(7):668李宁,范雪梅,王义明,等.中南药学,2014,12(7):668
[5] Li G H,Li X,Ding X Z,et al. Shaanxi Medical Journal,2012,41(3):298李桂华,李潇,丁希正,等.陕西医学杂志,2012,41(3):298
[6] Xie P Y,Zhang Y,Wang X B,et al. Food Funct,2017,8(5):2000
[7] Dong Y,Yu Z M,Li H Y,et al. Chinese Journal of Chromato graphy,2016,34(8):811董宇,俞忠明,李洪玉,等.色谱,2016,34(8):811
[8] Hu R,Li D L. Northw est Pharmaceutical Journal,2008,23(2):118胡锐,李定莉.西北药学杂志,2008,23(2):118
[9] Zhao X,Zhang Y,Meng X,et al. J Chromatogr B Analyt Techno l Bio m ed Life Sci,2008,873(2):151
[10] Meng X S,Jiang M,Luo G A,et al. Liaoning Journal of Traditio nal Chinese Medicine,2012,39(2):218孟宪生,姜民,罗国安,等.辽宁中医杂志,2012,39(2):218
[11] Huang L Q. Liaoning Journal of Traditional Chinese Medicine,2000,27(10):469黄力强.辽宁中医杂志,2000,27(10):469
[12] Mangin E L Jr,Kugiyama K,Nguy J H,et al. Circ Res,1993,72(1):161
[13] Engelmann B,Zieseniss S,Brand K,et al. Arterioscler Thro m b Vasc Bio l,1999,19(1):47
[14] Huang M Y,Tang Y P,Li W X,et al. China Journal of Chinese Materia Medica,2013,38(20):3576黄美艳,唐于平,李伟霞,等.中国中药杂志,2013,38(20):3576
[15] Li X P,Zhao S P,Gao M. Medical Research New sletter,2004,33(4):30李向平,赵水平,高梅.医学研究通讯,2004,33(4):30
[16] Li W X.[PhD Dissertation]. Nanjing:Nanjing University of Traditio nal Chinese Medicine,2014李伟霞.[博士学位论文].南京:南京中医药大学,2014
[17] Yuan Z W,Zhong L J,Ji P,et al. Natural Product Research and Develo pm ent,2018,30(4):559袁子文,钟立甲,纪鹏,等.天然产物研究与开发,2018,30(4):559
[18] Zheng J H,Chen J J,Jian W X,et al. Lishizhen Medicine and Materia Medica Research,2015,26(2):485郑景辉,陈建军,简维雄,等.时珍国医国药,2015,26(2):485
[2] Li W X, Huang M Y, Tang Y P, et al. Chinese Pharm aco lo gical Bulletin,2011,27(12):1761李伟霞,黄美艳,唐于平,等.中国药理学通报,2011,27(12):1761
[3] Wang C,Jia X Y,Hou L W,et al. Acta Pharmaceutica Sinica,2018,53(6):980王翠,贾雪洋,侯璐文,等.药学学报,2018,53(6):980
[4] Li N,Fan X M,Wang Y M,et al. Central South Pharmacy,2014,12(7):668李宁,范雪梅,王义明,等.中南药学,2014,12(7):668
[5] Li G H,Li X,Ding X Z,et al. Shaanxi Medical Journal,2012,41(3):298李桂华,李潇,丁希正,等.陕西医学杂志,2012,41(3):298
[6] Xie P Y,Zhang Y,Wang X B,et al. Food Funct,2017,8(5):2000
[7] Dong Y,Yu Z M,Li H Y,et al. Chinese Journal of Chromato graphy,2016,34(8):811董宇,俞忠明,李洪玉,等.色谱,2016,34(8):811
[8] Hu R,Li D L. Northw est Pharmaceutical Journal,2008,23(2):118胡锐,李定莉.西北药学杂志,2008,23(2):118
[9] Zhao X,Zhang Y,Meng X,et al. J Chromatogr B Analyt Techno l Bio m ed Life Sci,2008,873(2):151
[10] Meng X S,Jiang M,Luo G A,et al. Liaoning Journal of Traditio nal Chinese Medicine,2012,39(2):218孟宪生,姜民,罗国安,等.辽宁中医杂志,2012,39(2):218
[11] Huang L Q. Liaoning Journal of Traditional Chinese Medicine,2000,27(10):469黄力强.辽宁中医杂志,2000,27(10):469
[12] Mangin E L Jr,Kugiyama K,Nguy J H,et al. Circ Res,1993,72(1):161
[13] Engelmann B,Zieseniss S,Brand K,et al. Arterioscler Thro m b Vasc Bio l,1999,19(1):47
[14] Huang M Y,Tang Y P,Li W X,et al. China Journal of Chinese Materia Medica,2013,38(20):3576黄美艳,唐于平,李伟霞,等.中国中药杂志,2013,38(20):3576
[15] Li X P,Zhao S P,Gao M. Medical Research New sletter,2004,33(4):30李向平,赵水平,高梅.医学研究通讯,2004,33(4):30
[16] Li W X.[PhD Dissertation]. Nanjing:Nanjing University of Traditio nal Chinese Medicine,2014李伟霞.[博士学位论文].南京:南京中医药大学,2014
[17] Yuan Z W,Zhong L J,Ji P,et al. Natural Product Research and Develo pm ent,2018,30(4):559袁子文,钟立甲,纪鹏,等.天然产物研究与开发,2018,30(4):559
[18] Zheng J H,Chen J J,Jian W X,et al. Lishizhen Medicine and Materia Medica Research,2015,26(2):485郑景辉,陈建军,简维雄,等.时珍国医国药,2015,26(2):485