Abstract
Despite the high prevalence of alcoholic liver disease, its identification and characterization remain poor, especially in early stages such as alcoholic fatty liver disease and alcoholic steatohepatitis. This latter implies diagnostic difficulties, few therapeutic options and unclear mechanisms of action. To elucidate the metabolic alterations and pinpoint affected biochemical pathways, alcoholic steatohepatitis was simulated in vitro by exposing HepaRG cells to ethanol (IC10, 368 mM) and tumor necrosis factor alpha (TNF-α, 50 ng/mL) for 24 h. This combined exposure was compared to solely ethanol-exposed as well as -nonexposed cells. Four different metabolomics platforms were used combining liquid chromatography, high-resolution mass spectrometry and drift tube ion mobility to elucidate both intracellular and extracellular metabolic alterations. Some of the key findings include the influence of TNF-α in the upregulation of hepatic triglycerides and the downregulation of hepatic phosphatidylethanolamines and phosphatidylcholines. S-Adenosylmethionine showed to play a central role in the progression of alcoholic steatohepatitis. In addition, fatty acyl esters of hydroxy fatty acid (FAHFA)-containing triglycerides were detected for the first time in human hepatocytes and their alterations showed a potentially important role during the progression of alcoholic steatohepatitis. Ethoxylated phosphorylcholine was identified as a potential new biomarker of ethanol exposure.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability statement
Raw datafiles are available through the MassIVE repository (https://massive.ucsd.edu/ProteoSAFe/) with the data set identifier MSV000090773.
References
Aagaard NK, Thøgersen T, Grøfte T, Greisen J, Vilstrup H (2004) Alcohol acutely down-regulates urea synthesis in normal men. Alcoholism Clin Exp Res 28(5):697–701. https://doi.org/10.1097/01.ALC.0000125355.31808.DC
Avila MA, García-Trevijano ER, Lu SC, Corrales FJ, Mato JM (2004) Methylthioadenosine. Int J Biochem Cell Biol 36(11):2125–2130. https://doi.org/10.1016/J.BIOCEL.2003.11.016
Aw TY, Jones DP (1983) Intracellular inhibition of UDP-glucose dehydrogenase during ethanol oxidation. Chem Biol Interact 43(3):283–288. https://doi.org/10.1016/0009-2797(83)90112-6
Bailey SM, Patel VB, Young TA, Asayama K, Cunningham CC (2001) Chronic ethanol consumption alters the glutathione/glutathione peroxidase-1 system and protein oxidation status in rat liver. Alcoholism Clin Exp Res 25(5):726–733. https://doi.org/10.1111/J.1530-0277.2001.TB02273.X
Baykara B, Mıcılı SC, Tugyan K, Tekmen I, Bagriyanik H, Sonmez U, Sonmez A, Oktay G, Yener N, Ozbal S (2012) The protective effects of carnosine in alcohol-induced hepatic injury in rats. Toxicol Ind Health 30(1):25–32. https://doi.org/10.1177/0748233712446722
Beatriz MG, Morales JM, Rodrigo JM, Del Olmo J, Serra MA, Ferrández A, Celda B, Monleón D (2011) Metabolic profile of chronic liver disease by NMR spectroscopy of human biopsies. Int J Mol Med 27(1):111–117. https://doi.org/10.3892/IJMM.2010.563/HTML
Beirnaert C, Cuykx M, Bijtebier S (2019) MetaboMeeseeks: helper functions for metabolomics analysis. https://github.com/Beirnaert/MetaboMeeseeks/. Accessed 2 Dec 2022
Bilbao A, Gibbons BC, Stow SM, Kyle JE, Bloodsworth KJ, Payne SH, Smith RD, Ibrahim YM, Baker ES, Fjeldsted JC (2021) A preprocessing tool for enhanced ion mobility-mass spectrometry-based omics workflows. J Proteome Res 21(3):798–807. https://doi.org/10.1021/ACS.JPROTEOME.1C00425
Boeckmans J, Buyl K, Natale A, Vandenbempt V, Branson S, De Boe V, Rogiers V, De Kock J, Rodrigues RM, Vanhaecke T (2019) Elafibranor restricts lipogenic and inflammatory responses in a human skin stem cell-derived model of NASH. Pharmacol Res 144:377–389. https://doi.org/10.1016/J.PHRS.2019.04.016
Boeckmans J, Natale A, Rombaut M, Buyl K, Vanhaecke T, Rogiers V, Rodrigues RM, De Kock J (2020) Flow cytometric quantification of neutral lipids in a human skin stem cell-derived model of NASH. MethodsX 7:101068. https://doi.org/10.1016/J.MEX.2020.101068
Brusilow SW (1991) Phenylacetylglutamine may replace urea as a vehicle for waste nitrogen excretion. Pediatr Res 29(2):147–150. https://doi.org/10.1203/00006450-199102000-00009
Cahill A, Cunningham CC, Adachi M, Ishii H, Bailey SM, Fromenty B, Davies A (2002) Effects of alcohol and oxidative stress on liver pathology: the role of the mitochondrion. Alcoholism Clin Exp Res 26(6):907–915. https://doi.org/10.1111/J.1530-0277.2002.TB02621.X
Callans DJ, Wacker LS, Mitchell MC (1987) Effects of ethanol feeding and withdrawal on plasma glutathione elimination in the rat. Hepatology 7(3):496–501. https://doi.org/10.1002/HEP.1840070314
Calzada E, Onguka O, Claypool SM (2016) Phosphatidylethanolamine metabolism in health and disease. Int Rev Cell Mol Biol 321:29–88. https://doi.org/10.1016/BS.IRCMB.2015.10.001
Cederbaum AI (2010) Hepatoprotective effects of S-adenosyl-l-methionine against alcohol- and cytochrome P450 2E1-induced liver injury. World J Gastroenterol: WJG 16(11):1366–1376. https://doi.org/10.3748/WJG.V16.I11.1366
Clugston RD, Gao MA, Blaner WS (2017) The hepatic lipidome: a gateway to understanding the pathogenesis of alcohol-induced fatty liver. Curr Mol Pharmacol 10(3):195–206. https://doi.org/10.2174/1874467208666150817111419
da Silva KM, Iturrospe E, Heyrman J, Koelmel JP, Cuykx M, Vanhaecke T, Covaci A, van Nuijs ALN (2021) Optimization of a liquid chromatography-ion mobility-high resolution mass spectrometry platform for untargeted lipidomic and application to HepaRG cell extracts. Talanta 235:122808. https://doi.org/10.1016/j.talanta.2021.122808
Dean JM, Lodhi IJ (2018) Structural and functional roles of ether lipids. Protein Cell 9(2):196–206. https://doi.org/10.1007/S13238-017-0423-5
DeFelice BC, Mehta SS, Samra S, Čajka T, Wancewicz B, Fahrmann JF, Fiehn O (2017) Mass spectral feature list optimizer (MS-FLO): a tool to minimize false positive peak reports in untargeted liquid chromatography–mass spectroscopy (LC–MS) data processing. Anal Chem 89(6):3250–3255. https://doi.org/10.1021/ACS.ANALCHEM.6B04372
Dif N, Euthine V, Gonnet E, Laville M, Vidal H, Lefai E (2006) Insulin activates human sterol-regulatory-element-binding protein-1c (SREBP-1c) promoter through SRE motifs. Biochem J 400(Pt 1):179–188. https://doi.org/10.1042/BJ20060499
Endo M, Masaki T, Seike M, Yoshimatsu H (2007) TNF-α induces hepatic steatosis in mice by enhancing gene expression of sterol regulatory element binding protein-1c (SREBP-1c). Exp Biol Med 232(5):614–621. https://doi.org/10.3181/00379727-232-2320614
European Association for the Study of the Liver (2018) EASL clinical practice guidelines: management of alcohol-related liver disease. J Hepatol 69(1):154–181. https://doi.org/10.1016/J.JHEP.2018.03.018
European Medicines Agency EMA (2011) Guideline on bioanalytical method validation. https://www.ema.europa.eu. Accessed 10 Nov 2022
Fernández-Checa JC, Kaplowitz N, García-Ruiz C, Colell A, Miranda M, Marí M, Ardite E, Morales A (1997) GSH transport in mitochondria: defense against TNF-induced oxidative stress and alcohol-induced defect. Am J Physiol Gastrointest Liver Physiol 273(1):G7–G17. https://doi.org/10.1152/AJPGI.1997.273.1.G7
Gaude E, Chignola F, Spiliotopoulos D, Spitaleri A, Ghitti M, Garcia-Manteiga MJ, Mari S, Musco G (2013) muma, An R package for metabolomics univariate and multivariate statistical analysis. Curr Metabol 1(2):180–189. https://doi.org/10.2174/2213235x11301020005
Glavind E, Aagaard NK, Grønbæk H, Møller HJ, Orntoft NW, Vilstrup H, Thomsen KL (2016) Alcoholic hepatitis markedly decreases the capacity for urea synthesis. PLoS ONE 11(7):e0158388. https://doi.org/10.1371/JOURNAL.PONE.0158388
Goracci L, Tortorella S, Tiberi P, Pellegrino RM, Di Veroli A, Valeri A, Cruciani G (2017) Lipostar, a comprehensive platform-neutral cheminformatics tool for lipidomics. Anal Chem 89(11):6257–6264. https://doi.org/10.1021/ACS.ANALCHEM.7B01259
Grunfeld C, Verdier JA, Neese R, Moser AH, Feingold KR (1988) Mechanisms by which tumor necrosis factor stimulates hepatic fatty acid synthesis in vivo. J Lipid Res 29:1327–1335. https://doi.org/10.1016/S0022-2275(20)38435-2
Guney Y, Ozel Turkcu U, Hicsonmez A, Nalca Andrieu M, Guney HZ, Bilgihan A, Kurtman C (2006) Carnosine may reduce lung injury caused by radiation therapy. Med Hypotheses 66(5):957–959. https://doi.org/10.1016/J.MEHY.2005.11.023
Hernández-Alvarez MI, Sebastián D, Vives S, Ivanova S, Bartoccioni P, Kakimoto P, Plana N, Veiga SR, Hernández V, Vasconcelos N, Peddinti G, Adrover A, Jové M, Pamplona R, Gordaliza-Alaguero I, Calvo E, Cabré N, Castro R, Kuzmanic A, Zorzano A (2019) Deficient endoplasmic reticulum-mitochondrial phosphatidylserine transfer causes liver disease. Cell 177(4):881.e17-895.e17. https://doi.org/10.1016/j.cell.2019.04.010
Holmuhamedov EL, Czerny C, Beeson CC, Lemasters JJ (2012) Ethanol suppresses ureagenesis in rat hepatocytes: role of acetaldehyde*. J Biol Chem 287(10):7692. https://doi.org/10.1074/JBC.M111.293399
Horai H, Arita M, Kanaya S, Nihei Y, Ikeda T, Suwa K, Ojima Y, Tanaka K, Tanaka S, Aoshima K, Oda Y, Kakazu Y, Kusano M, Tohge T, Matsuda F, Sawada Y, Hirai M, Nakanishi H, Ikeda K, Nishioka T (2010) MassBank: a public repository for sharing mass spectral data for life sciences. J Mass Spectrom JMS 45(7):703–714. https://doi.org/10.1002/JMS.1777
Hu M, Wang F, Li X, Rogers CQ, Liang X, Finck BN, Mitra MS, Zhang R, Mitchell DA, You M (2012) Regulation of hepatic lipin-1 by ethanol: role of AMPK-SREBP-1 signaling. Hepatology (baltimore, MD) 55(2):437–446. https://doi.org/10.1002/HEP.24708
Israelsen M, Kim M, Suvitaival T, Madsen BS, Hansen CD, Torp N, Trost K, Thiele M, Hansen T, Legido-Quigley C, Krag A (2021) Comprehensive lipidomics reveals phenotypic differences in hepatic lipid turnover in ALD and NAFLD during alcohol intoxication. JHEP Rep 3(5):100325. https://doi.org/10.1016/J.JHEPR.2021.100325
Iturrospe E, Da Silva KM, Talavera Andújar B, Cuykx M, Boeckmans J, Vanhaecke T, Covaci A, van Nuijs ALN (2021) An exploratory approach for an oriented development of an untargeted hydrophilic interaction liquid chromatography-mass spectrometry platform for polar metabolites in biological matrices. J Chromatogr A 1637:461807. https://doi.org/10.1016/j.chroma.2020.461807
Iturrospe E, Da Silva KM, Robeyns R, van de Lavoir M, Boeckmans J, Vanhaecke T, van Nuijs ALN, Covaci A (2022) Metabolic signature of ethanol-induced hepatotoxicity in HepaRG cells by LC–MS-based untargeted metabolomics. J Proteome Res. https://doi.org/10.1021/acs.jproteome.2c00029
Iturrospe E, da Silva KM, van de Lavoir M, Robeyns R, Cuykx M, Vanhaecke T, van Nuijs ALN, Covaci A (2023) Mass spectrometry-based untargeted metabolomics and lipidomics platforms to analyze cell culture extracts. Methods Mol Biol (clifton, NJ) 2571:189–206. https://doi.org/10.1007/978-1-0716-2699-3_19
Jankevics A, Lloyd GR, Weber RJM (2022) pmp: Peak matrix processing and signal batch correction for metabolomics datasets (R package version 1.10.0.). https://doi.org/10.18129/B9.bioc.pmp
Jaremek M, Yu Z, Mangino M, Mittelstrass K, Prehn C, Singmann P, Xu T, Dahmen N, Weinberger KM, Suhre K, Peters A, Döring A, Hauner H, Adamski J, Illig T, Spector TD, Wang-Sattler R (2013) Alcohol-induced metabolomic differences in humans. Transl Psychiatry 3(7):e276–e276. https://doi.org/10.1038/tp.2013.55
Jeon S, Carr R (2020) Alcohol effects on hepatic lipid metabolism. J Lipid Res 61(4):470–479. https://doi.org/10.1194/JLR.R119000547
Jewell SA, Di Monte D, Gentile A, Guglielmi A, Altomare E, Albano O (1986) Decreased hepatic glutathione in chronic alcoholic patients. J Hepatol 3(1):1–6. https://doi.org/10.1016/S0168-8278(86)80139-8
Kanety H, Feinstein R, Papa MZ, Hemi R, Karasik A (1995) Tumor necrosis factor alpha-induced phosphorylation of insulin receptor substrate-1 (IRS-1). Possible mechanism for suppression of insulin-stimulated tyrosine phosphorylation of IRS-1. J Biol Chem 270(40):23780–23784. https://doi.org/10.1074/JBC.270.40.23780
Kastl L, Sauer SW, Ruppert T, Beissbarth T, Becker MS, Süss D, Krammer PH, Gülow K (2014) TNF-α mediates mitochondrial uncoupling and enhances ROS-dependent cell migration via NF-κB activation in liver cells. FEBS Lett 588(1):175–183. https://doi.org/10.1016/J.FEBSLET.2013.11.033
Kawaratani H, Tsujimoto T, Douhara A, Takaya H, Moriya K, Namisaki T, Noguchi R, Yoshiji H, Fujimoto M, Fukui H (2013) The effect of inflammatory cytokines in alcoholic liver disease. Mediat Inflamm 2013:495156. https://doi.org/10.1155/2013/495156
Klåvus A, Kokla M, Noerman S, Koistinen VM, Tuomainen M, Zarei I, Meuronen T, Häkkinen MR, Rummukainen S, Babu AF, Sallinen T, Kärkkäinen O, Paananen J, Broadhurst D, Brunius C, Hanhineva K (2020) “Notame”: workflow for non-targeted LC–MS metabolic profiling. Metabolites 10(4):135. https://doi.org/10.3390/METABO10040135
Koelmel JP, Kroeger NM, Gill EL, Ulmer CZ, Bowden JA, Patterson RE, Yost RA, Garrett TJ (2017a) Expanding lipidome coverage using LC–MS/MS data-dependent acquisition with automated exclusion list generation. J Am Soc Mass Spectrom 28(5):908–917. https://doi.org/10.1007/s13361-017-1608-0
Koelmel JP, Kroeger NM, Ulmer CZ, Bowden JA, Patterson RE, Cochran JA, Beecher CWW, Garrett TJ, Yost RA (2017b) LipidMatch: an automated workflow for rule-based lipid identification using untargeted high-resolution tandem mass spectrometry data. BMC Bioinform 18(1):331. https://doi.org/10.1186/s12859-017-1744-3
Koelmel JP, Tan WY, Li Y, Bowden JA, Ahmadireskety A, Patt AC, Orlicky DJ, Mathé E, Kroeger NM, Thompson DC, Cochran JA, Golla JP, Kandyliari A, Chen Y, Charkoftaki G, Guingab-Cagmat JD, Tsugawa H, Arora A, Veselkov K, Vasilou V (2021) Lipidomics and redox lipidomics indicate early stage alcohol-induced liver damage. Hepatol Commun 6:513–525. https://doi.org/10.1002/HEP4.1825
Kuda O, Brezinova M, Silhavy J, Landa V, Zidek V, Dodia C, Kreuchwig F, Vrbacky M, Balas L, Durand T, Hübner N, Fisher AB, Kopecky J, Pravenec M (2018) Nrf2-mediated antioxidant defense and peroxiredoxin 6 are linked to biosynthesis of palmitic acid ester of 9-hydroxystearic acid. Diabetes 67(6):1190–1199. https://doi.org/10.2337/DB17-1087
Lange M, Angelidou G, Ni Z, Criscuolo A, Schiller J, Blüher M, Fedorova M (2021) AdipoAtlas: a reference lipidome for human white adipose tissue. Cell Rep Med 2(10):100407. https://doi.org/10.1016/J.XCRM.2021.100407
Lawler JF, Yin M, Diehl AM, Roberts E, Chatterjee S (1998) Tumor necrosis factor-α stimulates the maturation of sterol regulatory element binding protein-1 in human hepatocytes through the action of neutral sphingomyelinase. J Biol Chem 273(9):5053–5059. https://doi.org/10.1074/jbc.273.9.5053
Li YQ, Prentice DA, Howard ML, Mashford ML, Wilson JS, Desmond PV (2000) Alcohol up-regulates udp-glucuronosyltransferase mrna expression in rat liver and in primary rat hepatocyte culture. Life Sci 66(7):575–584. https://doi.org/10.1016/S0024-3205(99)00630-X
Lieber CS, Robins SJ, Leo MA (1994) Hepatic phosphatidylethanolamine methyltransferase activity is decreased by ethanol and increased by phosphatidylcholine. Alcohol Clin Exp Res 18(3):592–595. https://doi.org/10.1111/J.1530-0277.1994.TB00915.X
Männistö V, Kaminska D, Kärjä V, Tiainen M, de Mello VD, Hanhineva K, Soininen P, Ala-Korpela M, Pihlajamäki J (2019) Total liver phosphatidylcholine content associates with non-alcoholic steatohepatitis and glycine N-methyltransferase expression. Liver Int 39(10):1895–1905. https://doi.org/10.1111/LIV.14174
Maudens KE, Patteet L, van Nuijs ALN, Van Broekhoven C, Covaci A, Neels H (2014) The influence of the body mass index (BMI) on the volume of distribution of ethanol. Forensic Sci Int 243:74–78. https://doi.org/10.1016/J.FORSCIINT.2014.04.036
McLean S, Davies NW, Nichols DS, McLeod BJ (2015) Triacylglycerol estolides, a new class of mammalian lipids, in the paracloacal gland of the Brushtail Possum (Trichosurus vulpecula). Lipids 50(6):591–604. https://doi.org/10.1007/S11745-015-4025-9
Meikle PJ, Mundra PA, Wong G, Rahman K, Huynh K, Barlow CK, Duly AMP, Haber PS, Whitfield JB, Seth D (2015) Circulating lipids are associated with alcoholic liver cirrhosis and represent potential biomarkers for risk assessment. PLoS ONE 10(6):e0130346. https://doi.org/10.1371/JOURNAL.PONE.0130346
Murphy K, Viroli C, Gormley IC (2020) Infinite mixtures of infinite factor analysers. Bayesian Anal 15(3):937–963. https://doi.org/10.1214/19-BA1179
Nagy LE (2015) The role of innate immunity in alcoholic liver disease. Alcohol Res Curr Rev 37(2):237–250 (/pmc/articles/PMC4590620/)
Naz S, Vallejo M, García A, Barbas C (2014) Method validation strategies involved in non-targeted metabolomics. J Chromatogr A 1353:99–105. https://doi.org/10.1016/j.chroma.2014.04.071
Ni Z, Angelidou G, Lange M, Hoffmann R, Fedorova M (2017) LipidHunter identifies phospholipids by high-throughput processing of LC–MS and shotgun lipidomics datasets. Anal Chem 89(17):8800–8807. https://doi.org/10.1021/ACS.ANALCHEM.7B01126
Pi J, Wu X, Feng Y (2016) Fragmentation patterns of five types of phospholipids by ultra-high-performance liquid chromatography electrospray ionization quadrupole time-of-flight tandem mass spectrometry. Anal Methods 8(6):1319–1332. https://doi.org/10.1039/C5AY00776C
Picache JA, Rose BS, Balinski A, Leaptrot KL, Sherrod SD, May JC, McLean JA (2019) Collision cross section compendium to annotate and predict multi-omic compound identities. Chem Sci 10(4):983–993. https://doi.org/10.1039/C8SC04396E
Popa C, Netea MG, Van Riel PLCM, Van Der Meer JWM, Stalenhoef AFH (2007) The role of TNF-α in chronic inflammatory conditions, intermediary metabolism, and cardiovascular risk. J Lipid Res 48(4):751–762. https://doi.org/10.1194/JLR.R600021-JLR200
Puri P, Baillie RA, Wiest MM, Mirshahi F, Choudhury J, Cheung O, Sargeant C, Contos MJ, Sanyal AJ (2007) A lipidomic analysis of nonalcoholic fatty liver disease. Hepatology 46(4):1081–1090. https://doi.org/10.1002/HEP.21763
Puri P, Xu J, Vihervaara T, Katainen R, Ekroos K, Daita K, Min HK, Joyce A, Mirshahi F, Tsukamoto H, Sanyal AJ (2016) Alcohol produces distinct hepatic lipidome and eicosanoid signature in lean and obese. J Lipid Res 57(6):1017–1028. https://doi.org/10.1194/JLR.M066175
Raina N, Matsui J, Cunnane SC, Jeejeebhoy KN (1995) Effect of tumor necrosis factor-α on triglyceride and phospholipid content and fatty acid composition of liver and carcass in rats. Lipids 30(8):713–718. https://doi.org/10.1007/BF02537797
Richardson TA, Sherman M, Kalman D, Morgan ET (2006) Expression of UDP-glucuronosyltransferase isoform mRNAs during inflammation and infection in mouse liver and kidney. Drug Metab Dispos Biol Fate Chem 34(3):351–353. https://doi.org/10.1124/DMD.105.007435
Robin MA, Demeilliers C, Sutton A, Paradis V, Maisonneuve C, Dubois S, Poirel O, Lettéron P, Pessayre D, Fromenty B (2005) Alcohol increases tumor necrosis factor α and decreases nuclear factor-κb to activate hepatic apoptosis in genetically obese mice. Hepatology 42(6):1280–1290. https://doi.org/10.1002/HEP.20949
Rodriguez DA, Moncada C, Núñez MT, Lavandero S, Ponnappa BC, Israel Y (2004) Ethanol increases tumor necrosis factor-alpha receptor-1 (TNF-R1) levels in hepatic, intestinal, and cardiac cells. Alcohol 33(1):9–15. https://doi.org/10.1016/J.ALCOHOL.2004.03.001
Ross DH, Cho JH, Xu L (2020) Breaking down structural diversity for comprehensive prediction of ion-neutral collision cross sections. Anal Chem 92(6):4548–4557. https://doi.org/10.1021/ACS.ANALCHEM.9B05772
Saccenti E, Hoefsloot HCJ, Smilde AK, Westerhuis JA, Hendriks MMWB (2014) Reflections on univariate and multivariate analysis of metabolomics data. Metabolomics 10(3):361–374. https://doi.org/10.1007/s11306-013-0598-6
Sakhuja P (2014) Pathology of alcoholic liver disease, can it be differentiated from nonalcoholic steatohepatitis? World J Gastroenterol WJG 20(44):16474–16479. https://doi.org/10.3748/WJG.V20.I44.16474
Sanchez-Dominguez CN, Gallardo-Blanco HL, Salinas-Santander MA, Ortiz-Lopez R (2018) Uridine 5ʹ-diphospho-glucronosyltrasferase: its role in pharmacogenomics and human disease (review). Exp Ther Med 16(1):3–11. https://doi.org/10.3892/ETM.2018.6184
Saponara E, Penno C, Orsini V, Wang Z-Y, Fischer A, Aebi A, Matadamas-Guzman ML, Brun V, Fischer B, Brousseau M, O’donnell P, Turner J, Meyer AG, Bollepalli L, D’ario G, Roma G, Carbone W, Annunziato S, Obrecht M (2022) Loss of hepatic leucine-rich repeat-containing G-protein–coupled receptors 4 and 5 promotes nonalcoholic fatty liver disease. Am J Pathol. https://doi.org/10.1016/J.AJPATH.2022.10.00
Schymanski EL, Jeon J, Gulde R, Fenner K, Ruff M, Singer HP, Hollender J (2014) Identifying small molecules via high resolution mass spectrometry: communicating confidence. Environ Sci Technol 48(4):2097–2098. https://doi.org/10.1021/es5002105
Seitz HK, Bataller R, Cortez-Pinto H, Gao B, Gual A, Lackner C, Mathurin P, Mueller S, Szabo G, Tsukamoto H (2018) Alcoholic liver disease. Nat Rev Dis Primers 4(1):16. https://doi.org/10.1038/S41572-018-0014-7
Seo W, Jeong WI (2016) Hepatic non-parenchymal cells: master regulators of alcoholic liver disease? World J Gastroenterol 22(4):1348–1356. https://doi.org/10.3748/WJG.V22.I4.1348
Shi C, Wang L, Zhou K, Shao M, Lu Y, Wu T (2020) Targeted metabolomics identifies differential serum and liver amino acids biomarkers in rats with alcoholic liver disease. J Nutr Sci Vitaminol 66(6):536–544. https://doi.org/10.3177/JNSV.66.536
Shim YR, Jeong WI (2020) Recent advances of sterile inflammation and inter-organ cross-talk in alcoholic liver disease. Exp Mol Med 52(5):772–780. https://doi.org/10.1038/s12276-020-0438-5
Stickel F, Seitz H (2003) Ethanol and methyl transfer. In: Watson RR, Preedy VR (eds) Nutrition and alcohol. CRC Press, Boca Raton, pp 57–71. https://doi.org/10.1201/9780203507636.CH4
Su Jung K, Su Hee K, Ji Hyun K, Shin H, Hyun Ju Y (2016) Understanding metabolomics in biomedical research. Endocrinol Metab 31(1):7–16. https://doi.org/10.3803/ENM.2016.31.1.7
Sun X, Ye C, Deng Q, Chen J, Guo C (2021) Contribution of glutaredoxin-1 to Fas s-glutathionylation and inflammation in ethanol-induced liver injury. Life Sci 264:118678. https://doi.org/10.1016/J.LFS.2020.118678
Tacer KF, Kuzman D, Seliškar M, Pompon D, Rozman D (2007) TNF-α interferes with lipid homeostasis and activates acute and proatherogenic processes. Physiol Genomics 31(2):216–227. https://doi.org/10.1152/PHYSIOLGENOMICS.00264.2006
Tan D, Ertunc ME, Konduri S, Zhang J, Pinto AM, Chu Q, Kahn BB, Siegel D, Saghatelian A (2019) Discovery of FAHFA-containing triacylglycerols and their metabolic regulation. J Am Chem Soc 141(22):8798–8806. https://doi.org/10.1021/JACS.9B00045
Thévenot EA, Roux A, Xu Y, Ezan E, Junot C (2015) Analysis of the human adult urinary metabolome variations with age, body mass index, and gender by implementing a comprehensive workflow for univariate and OPLS statistical analyses. J Proteome Res 14(8):3322–3335. https://doi.org/10.1021/acs.jproteome.5b00354
Tsugawa H, Kind T, Nakabayashi R, Yukihira D, Tanaka W, Cajka T, Saito K, Fiehn O, Arita M (2016) Hydrogen rearrangement rules: computational MS/MS fragmentation and structure elucidation using MS-FINDER software. Anal Chem 88(16):7946–7958. https://doi.org/10.1021/ACS.ANALCHEM.6B00770
Tsugawa H, Ikeda K, Takahashi M, Satoh A, Mori Y, Uchino H, Okahashi N, Yamada Y, Tada I, Bonini P, Higashi Y, Okazaki Y, Zhou Z, Zhu Z-J, Koelmel J, Cajka T, Fiehn O, Saito K, Arita M, Arita M (2020) A lipidome atlas in MS-DIAL 4. Nat Biotechnol 38(10):1159–1163. https://doi.org/10.1038/s41587-020-0531-2
Ventura R, Martínez-Ruiz I, Hernández-Alvarez MI (2022) Phospholipid membrane transport and associated diseases. Biomedicines 10(5):1201. https://doi.org/10.3390/biomedicines10051201
Vina J, Estrela JM, Guerri C, Romero FJ (1980) Effect of ethanol on glutathione concentration in isolated hepatocytes. Biochem J 188(2):549–552. https://doi.org/10.1042/BJ1880549
Vonghia L, Michielsen P, Dom G, Francque S (2014) Diagnostic challenges in alcohol use disorder and alcoholic liver disease. World J Gastroenterol WJG 20(25):8024–8032. https://doi.org/10.3748/WJG.V20.I25.8024
Wang Z, Yao T, Song Z (2010) Involvement and mechanism of DGAT2 upregulation in the pathogenesis of alcoholic fatty liver disease. J Lipid Res 51(11):3158–3165. https://doi.org/10.1194/JLR.M007948
Wu G, Yang J, Lv H, Jing W, Zhou J, Feng Y, Lin S, Yang Q, Hu J (2018) Taurine prevents ethanol-induced apoptosis mediated by mitochondrial or death receptor pathways in liver cells. Amino Acids 50(7):863–875. https://doi.org/10.1007/S00726-018-2561-3
Yin M, Wheeler MD, Kono H, Bradford BU, Gallucci RM, Luster MI, Thurman RG (1999) Essential role of tumor necrosis factor α in alcohol-induced liver injury in mice. Gastroenterology 117(4):942–952. https://doi.org/10.1016/S0016-5085(99)70354-9
Yore MM, Syed I, Moraes-Vieira PM, Zhang T, Herman MA, Homan EA, Patel RT, Lee J, Chen S, Peroni OD, Dhaneshwar AS, Hammarstedt A, Smith U, McGraw TE, Saghatelian A, Kahn BB (2014) Discovery of a class of endogenous mammalian lipids with anti-diabetic and anti-inflammatory effects. Cell 159(2):318–332. https://doi.org/10.1016/j.cell.2014.09.035
Zhou Z, Luo M, Chen X, Yin Y, Xiong X, Wang R, Zhu ZJ (2020) Ion mobility collision cross-section atlas for known and unknown metabolite annotation in untargeted metabolomics. Nat Commun 11(1):1–13. https://doi.org/10.1038/s41467-020-18171-8
Funding
Elias Iturrospe and Maria van de Lavoir acknowledge funding of the Research Scientific Foundation-Flanders (FWO)—project numbers 1161620N and 1120623N, respectively. Rani Robeyns and Katyeny Manuela da Silva were funded by the University of Antwerp (BOF-GOA—PS ID 41667 and BOF DOCPRO 4—Antigoon ID 36893, respectively). Further funding of this research has been provided by the University of Antwerp (UA, Belgium) and the Vrije Universiteit Brussel (VUB, Belgium).
Author information
Authors and Affiliations
Contributions
EI: conceptualization, methodology, investigation, formal analysis, validation, visualization, writing—original draft, writing—review and editing. RR: methodology, investigation, writing—review and editing. KMdS: methodology, writing—review and editing. MvdL: methodology, writing—review and editing. JB: methodology, writing—review and editing. TV: supervision, conceptualization, writing—review and editing, resources. ALNvN: supervision, conceptualization, writing—review and editing, resources. AC: supervision, conceptualization, writing—review and editing, resources.
Corresponding authors
Ethics declarations
Conflict of interest
The authors have no competing interests to declare that are relevant to the content of this article.
Ethical approval
Ethical approval for the use of HepaRG cells was provided by the Medical Ethics Committee of the University Hospital Brussels (reference number 143201941214).
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Iturrospe, E., Robeyns, R., da Silva, K.M. et al. Metabolic signature of HepaRG cells exposed to ethanol and tumor necrosis factor alpha to study alcoholic steatohepatitis by LC–MS-based untargeted metabolomics. Arch Toxicol 97, 1335–1353 (2023). https://doi.org/10.1007/s00204-023-03470-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-023-03470-y