液相色谱-质谱研究高尿酸血症大鼠血清代谢组学
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摘要
目的利用液相色谱-串联质谱技术,研究高尿酸血症(HUA)大鼠整体代谢谱的改变,筛选与高尿酸血症相关的潜在生物标志物。方法通过腺嘌呤和氧嗪酸钾灌胃3周,同时自由进食含10%酵母膏饲料方法建立高尿酸血症大鼠模型。采用基于超高效液相色谱-质谱(UPLC-MS/MS)的代谢组学方法,运用主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)分析比较模型组与正常大鼠血清的代谢谱差异。结果与对照组大鼠相比,在高尿酸模型组大鼠中发现并鉴定出14种潜在生物标志物,分别为尿酸、次黄嘌呤、尿囊素、肌酐、马尿酸、犬尿氨酸、色氨酸、硫酸吲哚酚、硫酸对甲酚、牛磺酸、棕榈酸、硬脂酸、溶血磷脂酰胆碱(LPC)(17∶0)和LPC(18∶0)。提示,高尿酸血症影响大鼠嘌呤代谢、氨基酸代谢、胆汁酸代谢、脂肪酸代谢及菌群代谢。结论本研究筛选出高尿酸模型大鼠血清中的14种差异代谢物,有助于解释高尿酸血症引起的代谢改变,可望为高尿酸血症的早期筛查、诊断和治疗提供帮助。
Objective To investigate the comprehensive serum metabolic profiles of hyperuricemia(HUA) rats and to identify potential biomarkers of hyperuricemia by liquid chromatography-tandem mass spectrometry. MethodsHyperuricemia rat model was established by consistent intragastric administration of adenine and potassium oxazinate for 3 weeks, rats were fed with 10% yeast extract at the same time. The metabolomics method based on ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS/MS), principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were used to find the difference of serum metabolic profiles between hyperuricemia model and control groups. Results Comparing with control rats, 14 potential biomarkers were identified in hyperuricemia rats, including uric acid, hypoxanthine, allantoin, creatinine, hippuric acid, kynurenine, tryptophan, indoxyl sulfate, p-cresol sulphate, taurine, palmitic acid, stearic acid, lyso-phosphatidylcholine(LPC)(17∶0) and LPC(18∶0). The results showed that hyperuricemia affected purine metabolism, amino acid metabolism, bile acid metabolism, fatty acid metabolism and gut microbiota metabolism in rats. Conclusions Fourteen different serum metabolites of hyperuricemia model rats were examined,which were helpful to explain the metabolic profiles changes caused by hyperuricemia,and to facilitate early screening,diagnosis and treatment of hyperuricemia.
Objective To investigate the comprehensive serum metabolic profiles of hyperuricemia(HUA) rats and to identify potential biomarkers of hyperuricemia by liquid chromatography-tandem mass spectrometry. MethodsHyperuricemia rat model was established by consistent intragastric administration of adenine and potassium oxazinate for 3 weeks, rats were fed with 10% yeast extract at the same time. The metabolomics method based on ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS/MS), principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were used to find the difference of serum metabolic profiles between hyperuricemia model and control groups. Results Comparing with control rats, 14 potential biomarkers were identified in hyperuricemia rats, including uric acid, hypoxanthine, allantoin, creatinine, hippuric acid, kynurenine, tryptophan, indoxyl sulfate, p-cresol sulphate, taurine, palmitic acid, stearic acid, lyso-phosphatidylcholine(LPC)(17∶0) and LPC(18∶0). The results showed that hyperuricemia affected purine metabolism, amino acid metabolism, bile acid metabolism, fatty acid metabolism and gut microbiota metabolism in rats. Conclusions Fourteen different serum metabolites of hyperuricemia model rats were examined,which were helpful to explain the metabolic profiles changes caused by hyperuricemia,and to facilitate early screening,diagnosis and treatment of hyperuricemia.
引文
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[4] Luan H, Wang X, Cai Z. Mass spectrometry-based metabolomics: Targeting the crosstalk between gut microbiota and brain in neurodegenerative disorders[J]. Mass Spectrom Rev, 2017: 1-12.
[5] Dankers AC, Mutsaers HA, Dijkman HB, et al. Hyperuricemia influences tryptophan metabolism via inhibition of multidrug resistance protein 4 (MRP4) and breast cancer resistance protein (BCRP)[J]. Biochim Biophys Acta, 2013, 1832: 1715-1722.
[6] Dou L, Bertrand E, Cerini C, et al. The uremic solutes p-cresol and indoxyl sulfate inhibit endothelial prolifera-tion and wound repair[J]. Kidney Int, 2004, 65: 442-451.
[7] Calvani R, Miccheli A, Capuani G, et al. Gut micro-biome-derived metabolites characterize a peculiar obese urinary metabotype[J]. Int J Obes (Lond), 2010, 34: 1095-1098.
[8] Richette P, Poitou C, Manivet P, et al. Weight Loss, xanthine oxidase, and serum urate levels: A prospective longitudinal study of obese patients[J]. Arthritis Care Res (Hoboken), 2016, 68: 1036-1042.
[9] Kim TH, Lee SS, Yoo JH, et al. The relationship between the regional abdominal adipose tissue distribution and the serum uric acid levels in people with type 2 diabetes mellitus[J]. Diabetol Metab Syndr, 2012, 4: 3-4.
[10] Forouhi NG, Koulman A, Sharp SJ, et al. Differences in the prospective association between individual plasma phospholipid saturated fatty acids and incident type 2 diabetes: the EPIC-InterAct case-cohort study[J]. The Lancet Diabetes Endo, 2014, 2: 810-818.
[11] 张淑丽, 王怡萍, 杜振霞, 等. 基于UPC～2-Q/TOF-MS技术的果糖诱导高尿酸血症大鼠血清脂质代谢组学研究[J]. 中国中药杂志, 2016: 1135-1139.
[12] Xu Q, Zhang M, Abeysekera IR, et al. High serum uric acid levels may increase mortality and major adverse cardiovascular events in patients with acute myocardial infarction[J]. Saudi Med J, 2017, 38: 577-585.
[2] 苗志敏, 赵世华, 王颜刚, 等. 山东沿海居民高尿酸血症及痛风的流行病学调查[J]. 中华内分泌代谢杂志, 2006: 421-425.
[3] Wang Z, Lin Y, Liu Y, et al. Serum uric acid levels and outcomes after acute ischemic stroke[J]. Mol Neurobiol, 2016, 53: 1753-1759.
[4] Luan H, Wang X, Cai Z. Mass spectrometry-based metabolomics: Targeting the crosstalk between gut microbiota and brain in neurodegenerative disorders[J]. Mass Spectrom Rev, 2017: 1-12.
[5] Dankers AC, Mutsaers HA, Dijkman HB, et al. Hyperuricemia influences tryptophan metabolism via inhibition of multidrug resistance protein 4 (MRP4) and breast cancer resistance protein (BCRP)[J]. Biochim Biophys Acta, 2013, 1832: 1715-1722.
[6] Dou L, Bertrand E, Cerini C, et al. The uremic solutes p-cresol and indoxyl sulfate inhibit endothelial prolifera-tion and wound repair[J]. Kidney Int, 2004, 65: 442-451.
[7] Calvani R, Miccheli A, Capuani G, et al. Gut micro-biome-derived metabolites characterize a peculiar obese urinary metabotype[J]. Int J Obes (Lond), 2010, 34: 1095-1098.
[8] Richette P, Poitou C, Manivet P, et al. Weight Loss, xanthine oxidase, and serum urate levels: A prospective longitudinal study of obese patients[J]. Arthritis Care Res (Hoboken), 2016, 68: 1036-1042.
[9] Kim TH, Lee SS, Yoo JH, et al. The relationship between the regional abdominal adipose tissue distribution and the serum uric acid levels in people with type 2 diabetes mellitus[J]. Diabetol Metab Syndr, 2012, 4: 3-4.
[10] Forouhi NG, Koulman A, Sharp SJ, et al. Differences in the prospective association between individual plasma phospholipid saturated fatty acids and incident type 2 diabetes: the EPIC-InterAct case-cohort study[J]. The Lancet Diabetes Endo, 2014, 2: 810-818.
[11] 张淑丽, 王怡萍, 杜振霞, 等. 基于UPC～2-Q/TOF-MS技术的果糖诱导高尿酸血症大鼠血清脂质代谢组学研究[J]. 中国中药杂志, 2016: 1135-1139.
[12] Xu Q, Zhang M, Abeysekera IR, et al. High serum uric acid levels may increase mortality and major adverse cardiovascular events in patients with acute myocardial infarction[J]. Saudi Med J, 2017, 38: 577-585.