基于UHPLC-MS的代谢组学技术研究婴儿巨细胞病毒肝炎湿热内蕴证（英文）
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摘要
目的利用代谢组学技术探讨人巨细胞病毒(HCMV)肝炎湿热内蕴证的证候实质。方法收集20例HCMV肝炎湿热内蕴证的患儿和20例正常对照组婴儿。采用超高效液相-线性离子阱/静电场轨道阱质谱联用技术对这些受试者的血浆及尿液样本进行代谢组学研究。通过正交偏最小二乘法对这些代谢数据进行分析,同时采用受试者工作曲线评价潜在生物标志物的特异性和敏感性。结果共检测到1 076个血浆代谢物和414个尿液代谢物。最终共鉴定出22个血浆代谢物和7个尿液代谢物,涉及鞘脂、甘油磷脂、组氨酸、甘油酯和脂肪酸代谢,其中鞘磷脂和甘油三酯可作为HCMV肝炎湿热内蕴证潜在的生物标记物。结论代谢物和生物标志物的确认为探讨HCMV肝炎湿热内蕴证的证候实质提供依据。
OBJECTIVE Using metabolomics to explore the essence of damp heat accumulation syndrome of infantile human cytomegalovirus hepatitis.METHODS 20 infantile HCMV hepatitis patients with damp heat accumulation syndrome and 20 healthy infants were recruited for investigation.Plasma and urine samples were analyzed using ultra-high-performance liquid chromatography-linear trap quadruple/orbitrap mass spectrometry(UHPLC-LTQ/Orbitrap-MS)to determine the alterations in metabolomic profiles of this syndrome.Orthogonal partial least squares-discriminate analysis(OPLS-DA)was applied to analyze the UHPLC-MS data obtained from these samples.The specificity and sensitivity of potential biomarkers were assessed according to the area under the receiver operator characteristic(ROC)curves.RESULTS The metabolomic analysis yielded1076 plasma compounds and 414 urine compounds.Among them,22 plasma and 7 urine differential metabolites relating to sphingolipid,glycerophospholipid,histidine,glycerolipid,and fatty acid metabolism were identified.Sphingomyelin and triglycerides were screened as potential biomarkers and showed excellent discriminant performance.CONCLUSION This research can provide insights into the essence of HCMV hepatitis damp heat accumulation syndrome.
OBJECTIVE Using metabolomics to explore the essence of damp heat accumulation syndrome of infantile human cytomegalovirus hepatitis.METHODS 20 infantile HCMV hepatitis patients with damp heat accumulation syndrome and 20 healthy infants were recruited for investigation.Plasma and urine samples were analyzed using ultra-high-performance liquid chromatography-linear trap quadruple/orbitrap mass spectrometry(UHPLC-LTQ/Orbitrap-MS)to determine the alterations in metabolomic profiles of this syndrome.Orthogonal partial least squares-discriminate analysis(OPLS-DA)was applied to analyze the UHPLC-MS data obtained from these samples.The specificity and sensitivity of potential biomarkers were assessed according to the area under the receiver operator characteristic(ROC)curves.RESULTS The metabolomic analysis yielded1076 plasma compounds and 414 urine compounds.Among them,22 plasma and 7 urine differential metabolites relating to sphingolipid,glycerophospholipid,histidine,glycerolipid,and fatty acid metabolism were identified.Sphingomyelin and triglycerides were screened as potential biomarkers and showed excellent discriminant performance.CONCLUSION This research can provide insights into the essence of HCMV hepatitis damp heat accumulation syndrome.
引文
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[2]BOPPANA SB,PASS RF,BRITT WJ,et al.Symptomatic congenital cytomegalovirus infection:neonatal morbidity and mortality[J].Pediatr Infect Dis J,1992,11(2):93-99.
[3]BOPPANA SB,ROSS SA,FOWLER KB,et al.Congenital cytomegalovirus infection:clinical outcome[J].Clin Infect Dis,2013,57:S178-S181.
[4]Dreher AM,ARORA N,FOWLER KB,et al.Spectrum of disease and outcome in children with symptomatic congenital cytomegalovirus infection[J].J Pediatr,2014,164(4):855-859.
[5]ZUPPAN CW,BUI HD,GRILL BG,et al.Diffuse hepatic fibrosis in congenital cytomegalovirus infection[J].J Pediatr Gastroenterol Nutr,1986,5(3):489-491.
[6]TEZER H,SEMEER G,KARA A,et al.Cytomegalovirus hepatitis and ganciclovir treatment in immunocompetent children[J].Turk J Pediatr,2008,50(3):228-234.
[7]BIRON KK.Antiviral drugs for cytomegalovirus diseases[J].Antiviral Res,2006,71(2/3):154-163.
[8]FIEHN O.Metabolomics-the link between genotypes and phenotypes[J].Plant Mol Biol,2002,48(1):155-171.
[9]GIKA HG,THEODORIDIS GA,PLUMB RS,et al.Current practice of liquid chromatography-mass spectrometry in metabolomics and metabonomics[J].J Pharm Biomed Anal,2014,87:12-25.
[10]WANG X,SUN H,ZHANG A,et al.Ultra-performance liquid chromatography coupled to mass spectrometry as a sensitive and powerful technology for metabolomic studies[J].J Sep Sci,2011,34:3451-3459.
[11]MUNGER J,BAJAD SU,COLLER HA,et al.Dynamics of the cellular metabolome during human cytomegalovirus infection[J].PLoS Pathog,2006,2(13):1165-1175.
[12]VASTAG L,KOYUNCU E,GRADY SL,et al.Divergent effects of human cytomegalovirus and herpes simplex virus-1on cellular metabolism[J].PLoS Pathog,2011,7:1-15.
[13]FANOS V,LOCCI E,NOTO A,et al.Urinary metabolomics in newborns infected by human cytomegalovirus:apreliminary investigation[J].Early Hum Dev,2013,89(S1):58-61.
[14]LIU Z,TIAN Y,WANG B,et al.Serum proteomics with SELDI-TOF-MS in congenital human cytomegalovirus hepatitis[J].J Med Virol,2007,79(10):1500-1505.
[15]BROADHURST D,KELL D.Statistical strategies for avoiding false discoveries in metabolomics and related experiments[J].Metabolomics,2006,2(4):171-196.
[16]MANICKLAL S,EMERY VC,LAZZARITTI T,et al.The"silent"global burden of congenital cytomegalovirus[J].Clin Microbiol Rev,2013,26(1):86-102.
[17]OZKAN TB,MISTIK R,DIKICI B,et al.Antiviral therapy in neonatal cholestatic cytomegalovirus hepatitis[J].BMC Gastroenterol,2007,7:9.
[18]PURDY JG,SHENK T,RABINOWITZ JD.Fatty acid elongase 7catalyzes lipidome remodeling essential for human cytomegalovirus replication[J].Cell Rep,2015,10(8):1375-1385.
[19]GRAMMATIKOS G.Persistence of HCV in acutely-infected patients depletes C24-ceramide and upregulates sphingosine and sphinganine serum levels[J].Int J Mol Sci,2016,17(6):922.
[20]DE ALMEIDA RF,FEDOROV A,PRIETO M.Sphingomyelin/phosphatidylcholine/cholesterol phase diagram:boundaries and composition of lipid rafts[J].Biophysic J,2003,85(4):2406-2416.
[21]TAFESSE FG,TAFESSE A,SUMANA S,et al.Intact sphingomyelin biosynthetic pathway is essential for intracellular transport of influenza virus glycoproteins[J].Proc Natl Acad Sci USA,2013,110(16):6406-6411.
[22]MARTIN-ACEBES MA,GABANDE-ROGRIGUEZ E,GARCIA-CABRERO AM,et al.et al.Host sphingomyelin increases West Nile virus infection in vivo[J].J Lipid Res,2016,57(3):422-432.
[23]HIRATA Y,IKEDA K,SUDOH M,et al.Self-enhancement of hepatitis C virus replication by promotion of specific sphingolipid biosynthesis[J].PLoS Pathog,2012,8(8):1-13.
[24]ZHENG SJ,QU F,LI JF,et al.Serum sphingomyelin has potential to reflect hepatic injury in chronic hepatitis B virus infection[J].Int J Infect Dis,2015,33:149-155.
[25]ALLAL C,BUISSON-BRENAC C,MARION V,et al.Human cytomegalovirus carries a cell-derived phospholipase A2required for infectivity[J].J Virol,2004,78:7717-7726.
[26]LIU A,KRAUSZ KW,FANG ZZ,et al.Gemfibrozil disrupts lysophosphatidylcholine and bile acid homeostasis via PPARalpha and its relevance to hepatotoxicity[J].Arch Toxicol,2014,88:983-996.
[27]LJUNGMAN P,GRIFFITHS P,PAYA C.Definitions of cytomegalovirus infection and disease in transplant recipients[J].Clin Infect Dis,2002,34:1094-1097.
[28]SCHENKEL LC,BAKOVIC M.Palmitic acid and oleic acid differentially regulate choline transporter-like 1levels and glycerolipid metabolism in skeletal muscle cells[J].Lipids,2014,49:731-744.