{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T19:13:53Z","timestamp":1775848433675,"version":"3.50.1"},"reference-count":97,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,5,19]],"date-time":"2021-05-19T00:00:00Z","timestamp":1621382400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia (FCT)","award":["PTDC\/BTM-SAL\/29297\/2017, POCI-01-0145-FEDER-029297, PTDC\/DTP-FTO\/2433\/2014, POCI-01-0145-FEDER-016659, PTDC\/BIA-MOL\/28607\/2017, POCI-01-0145-FEDER-028607, POCI-01-0145-FEDER-006980, NORTE-01-0145-FEDER-000028, PTDC\/ASP-HOR\/29152\/2017, POCI-01-0145-FEDER-0"],"award-info":[{"award-number":["PTDC\/BTM-SAL\/29297\/2017, POCI-01-0145-FEDER-029297, PTDC\/DTP-FTO\/2433\/2014, POCI-01-0145-FEDER-016659, PTDC\/BIA-MOL\/28607\/2017, POCI-01-0145-FEDER-028607, POCI-01-0145-FEDER-006980, NORTE-01-0145-FEDER-000028, PTDC\/ASP-HOR\/29152\/2017, POCI-01-0145-FEDER-0"]}]},{"DOI":"10.13039\/100010665","name":"H2020 Marie Sk\u0142odowska-Curie Actions","doi-asserted-by":"publisher","award":["722619, 734719"],"award-info":[{"award-number":["722619, 734719"]}],"id":[{"id":"10.13039\/100010665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004281","name":"Narodowe Centrum Nauki","doi-asserted-by":"publisher","award":["UMO-2018\/29\/B\/NZ1\/00589"],"award-info":[{"award-number":["UMO-2018\/29\/B\/NZ1\/00589"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Non-alcoholic steatohepatitis (NASH), one of the deleterious stages of non-alcoholic fatty liver disease, remains a significant cause of liver-related morbidity and mortality worldwide. In the current work, we used an exploratory data analysis to investigate time-dependent cellular and mitochondrial effects of different supra-physiological fatty acids (FA) overload strategies, in the presence or absence of fructose (F), on human hepatoma-derived HepG2 cells. We measured intracellular neutral lipid content and reactive oxygen species (ROS) levels, mitochondrial respiration and morphology, and caspases activity and cell death. FA-treatments induced a time-dependent increase in neutral lipid content, which was paralleled by an increase in ROS. Fructose, by itself, did not increase intracellular lipid content nor aggravated the effects of palmitic acid (PA) or free fatty acids mixture (FFA), although it led to an up-expression of hepatic fructokinase. Instead, F decreased mitochondrial phospholipid content, as well as OXPHOS subunits levels. Increased lipid accumulation and ROS in FA-treatments preceded mitochondrial dysfunction, comprising altered mitochondrial membrane potential (\u0394\u03a8m) and morphology, and decreased oxygen consumption rates, especially with PA. Consequently, supra-physiological PA alone or combined with F prompted the activation of caspase pathways leading to a time-dependent decrease in cell viability. Exploratory data analysis methods support this conclusion by clearly identifying the effects of FA treatments. In fact, unsupervised learning algorithms created homogeneous and cohesive clusters, with a clear separation between PA and FFA treated samples to identify a minimal subset of critical mitochondrial markers in order to attain a feasible model to predict cell death in NAFLD or for high throughput screening of possible therapeutic agents, with particular focus in measuring mitochondrial function.<\/jats:p>","DOI":"10.3390\/nu13051723","type":"journal-article","created":{"date-parts":[[2021,5,19]],"date-time":"2021-05-19T21:49:21Z","timestamp":1621460961000},"page":"1723","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells"],"prefix":"10.3390","volume":"13","author":[{"given":"Ricardo","family":"Amorim","sequence":"first","affiliation":[{"name":"CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, Portugal"},{"name":"CIQUP\/Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"},{"name":"PhD Programme in Experimental Biology and Biomedicine (PDBEB), Institute for Interdisciplinary Research (IIIUC), University of Coimbra, 3004-531 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9701-9431","authenticated-orcid":false,"given":"In\u00eas","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"Laboratory of Mitochondrial Biology and Metabolism, Nencki Institute of Experimental Biology of Polish Academy of Sciences, 02-093 Warsaw, Poland"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9355-2685","authenticated-orcid":false,"given":"Caroline","family":"Veloso","sequence":"additional","affiliation":[{"name":"CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, Portugal"}]},{"given":"Adriana","family":"Carvalho","sequence":"additional","affiliation":[{"name":"CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, Portugal"},{"name":"PhD Programme in Experimental Biology and Biomedicine (PDBEB), Institute for Interdisciplinary Research (IIIUC), University of Coimbra, 3004-531 Coimbra, Portugal"}]},{"given":"Rui","family":"Sim\u00f5es","sequence":"additional","affiliation":[{"name":"CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, Portugal"},{"name":"PhD Programme in Experimental Biology and Biomedicine (PDBEB), Institute for Interdisciplinary Research (IIIUC), University of Coimbra, 3004-531 Coimbra, Portugal"}]},{"given":"Francisco","family":"Pereira","sequence":"additional","affiliation":[{"name":"Center for Informatics and Systems, University of Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra, Portugal"},{"name":"Coimbra Polytechnic-ISEC, 3030-190 Coimbra, Portugal"}]},{"given":"Theresa","family":"Thiel","sequence":"additional","affiliation":[{"name":"Mediagnostic, D-72770 Reutlingen, Germany"}]},{"given":"Andrea","family":"Normann","sequence":"additional","affiliation":[{"name":"Mediagnostic, D-72770 Reutlingen, Germany"}]},{"given":"Catarina","family":"Morais","sequence":"additional","affiliation":[{"name":"Center for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Cal\u00e7ada Martim de Freitas, 3000-456 Coimbra, Portugal"}]},{"given":"Am\u00e1lia","family":"Jurado","sequence":"additional","affiliation":[{"name":"Center for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Cal\u00e7ada Martim de Freitas, 3000-456 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0789-4521","authenticated-orcid":false,"given":"Mariusz","family":"Wieckowski","sequence":"additional","affiliation":[{"name":"Laboratory of Mitochondrial Biology and Metabolism, Nencki Institute of Experimental Biology of Polish Academy of Sciences, 02-093 Warsaw, Poland"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0834-5698","authenticated-orcid":false,"given":"Jos\u00e9","family":"Teixeira","sequence":"additional","affiliation":[{"name":"CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, Portugal"},{"name":"CIQUP\/Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5201-9948","authenticated-orcid":false,"given":"Paulo","family":"Oliveira","sequence":"additional","affiliation":[{"name":"CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1097\/TP.0000000000002484","article-title":"Epidemiology of Nonalcoholic Fatty Liver Disease and Nonalcoholic Steatohepatitis: Implications for Liver Transplantation","volume":"103","author":"Younossi","year":"2019","journal-title":"Transplantation"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1038\/nrgastro.2013.41","article-title":"Progression of NAFLD to diabetes mellitus, cardiovascular disease or cirrhosis","volume":"10","author":"Anstee","year":"2013","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.fct.2014.01.027","article-title":"Multiple pathways are involved in palmitic acid-induced toxicity","volume":"67","author":"Park","year":"2014","journal-title":"Food Chem. Toxicol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"830","DOI":"10.1111\/j.1440-1746.2008.05733.x","article-title":"Differential effect of oleic and palmitic acid on lipid accumulation and apoptosis in cultured hepatocytes","volume":"24","author":"Ricchi","year":"2009","journal-title":"J. Gastroenterol. Hepatol."},{"key":"ref_5","first-page":"E275","article-title":"Saturated fatty acids induce endoplasmic reticulum stress and apoptosis independently of ceramide in liver cells","volume":"291","author":"Wei","year":"2006","journal-title":"Am. J. Physiol. Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1210\/en.2005-0570","article-title":"Saturated Fatty Acids Promote Endoplasmic Reticulum Stress and Liver Injury in Rats with Hepatic Steatosis","volume":"147","author":"Wang","year":"2006","journal-title":"Endocrinology"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"\u0160r\u00e1mek, J., N\u011bmcov\u00e1-F\u00fcrstov\u00e1, V., and Kov\u00e1\u0159, J. (2016). Kinase Signaling in Apoptosis Induced by Saturated Fatty Acids in Pancreatic \u03b2-Cells. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17091400"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1016\/j.molmet.2014.05.004","article-title":"ER calcium release promotes mitochondrial dysfunction and hepatic cell lipotoxicity in response to palmitate overload","volume":"3","author":"Egnatchik","year":"2014","journal-title":"Mol. Metab."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1495","DOI":"10.1002\/hep.22183","article-title":"The lysosomal-mitochondrial axis in free fatty acid-induced hepatic lipotoxicity","volume":"47","author":"Li","year":"2007","journal-title":"Hepatology"},{"key":"ref_10","first-page":"G236","article-title":"CHOP and AP-1 cooperatively mediate PUMA expression during lipoapoptosis","volume":"299","author":"Cazanave","year":"2010","journal-title":"Am. J. Physiol. Liver Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/j.metabol.2013.10.009","article-title":"Palmitate-induced Activation of Mitochondrial Metabolism Promotes Oxidative Stress and Apoptosis in H4IIEC3 Rat Hepatocytes","volume":"63","author":"Egnatchik","year":"2014","journal-title":"Metabolism"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"11663","DOI":"10.1074\/jbc.M115.649483","article-title":"Opposite Cross-Talk by Oleate and Palmitate on Insulin Signaling in Hepatocytes through Macrophage Activation","volume":"290","author":"Pardo","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1016\/j.jnutbio.2011.07.006","article-title":"Oleic acid activates peroxisome proliferator-activated receptor \u03b4 to compensate insulin resistance in steatotic cells","volume":"23","author":"Wu","year":"2012","journal-title":"J. Nutr. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Yang, J., Fern\u00e1ndez-Galilea, M., Mart\u00ednez-Fern\u00e1ndez, L., Gonz\u00e1lez-Muniesa, P., P\u00e9rez-Ch\u00e1vez, A., Mart\u00ednez, J.A., and Moreno-Aliaga, M.J. (2019). Oxidative Stress and Non-Alcoholic Fatty Liver Disease: Effects of Omega-3 Fatty Acid Supplementation. Nutrients, 11.","DOI":"10.3390\/nu11040872"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1186\/1476-511X-10-81","article-title":"\u03b1-Linolenic acid prevents endoplasmic reticulum stress-mediated apoptosis of stearic acid lipotoxicity on primary rat hepatocytes","volume":"10","author":"Zhang","year":"2011","journal-title":"Lipids Health Dis."},{"key":"ref_16","first-page":"1","article-title":"\u03c9-3 PUFAs ameliorate liver fibrosis and inhibit hepatic stellate cells proliferation and acti-vation by promoting YAP\/TAZ degradation","volume":"6","author":"Zhang","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Moreira, A.R.S., R\u00fcschenbaum, S., Schefczyk, S., Hendgen-Cotta, U., Rassaf, T., Broering, R., Hardtke-Wolenski, M., and Buitrago-Molina, L.E. (2020). 9-PAHSA Prevents Mitochondrial Dysfunction and Increases the Viability of Steatotic Hepatocytes. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21218279"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1097\/MCO.0b013e328361c4d1","article-title":"Carbohydrate intake and nonalcoholic fatty liver disease","volume":"16","year":"2013","journal-title":"Curr. Opin. Clin. Nutr. Metab. Care"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Agoun, H., Semiane, N., Mallek, A., Bellahreche, Z., Hammadi, S., Madjerab, M., Abdlalli, M., Khalkhal, A., and Dahmani, Y. (2019). High-carbohydrate diet-induced metabolic disorders in Gerbillus tarabuli (a new model of non-alcoholic fatty-liver disease). Protective effects of 20-hydroxyecdysone. Arch. Physiol. Biochem., 1\u20139.","DOI":"10.1080\/13813455.2019.1621350"},{"key":"ref_20","first-page":"G77","article-title":"Mechanisms of lysophosphatidylcholine-induced hepatocyte lipoapoptosis","volume":"302","author":"Kakisaka","year":"2012","journal-title":"Am. J. Physiol. Liver Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1074","DOI":"10.1111\/liv.12331","article-title":"Inhibition of ceramide de novo synthesis reduces liver lipid accumula-tion in rats with nonalcoholic fatty liver disease","volume":"34","author":"Kurek","year":"2014","journal-title":"Liver Int."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.tem.2012.04.005","article-title":"Role of ceramides in nonalcoholic fatty liver disease","volume":"23","author":"Pagadala","year":"2012","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1765","DOI":"10.1016\/j.bbadis.2015.05.015","article-title":"Recent insights on the role of cholesterol in non-alcoholic fatty liver disease","volume":"1852","author":"Arguello","year":"2015","journal-title":"Biochim. Biophys. Acta Mol. Basis Dis."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1128\/MCB.00420-13","article-title":"c-Jun N-Terminal Kinase 1\/c-Jun Activation of the p53\/MicroRNA 34a\/Sirtuin 1 Pathway Contributes to Apoptosis Induced by Deoxycholic Acid in Rat Liver","volume":"34","author":"Ferreira","year":"2014","journal-title":"Mol. Cell. Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"934","DOI":"10.1002\/hep.23797","article-title":"High-fructose, medium chain trans fat diet induces liver fibrosis and elevates plasma coenzyme Q9 in a novel murine model of obesity and nonalcoholic steatohepatitis","volume":"52","author":"Kohli","year":"2010","journal-title":"Hepatology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/j.diabet.2010.03.003","article-title":"Effects of short-term overfeeding with fructose, fat and fructose plus fat on plasma and hepatic lipids in healthy men","volume":"36","author":"Sobrecases","year":"2010","journal-title":"Diabetes Metab."},{"key":"ref_27","first-page":"284","article-title":"Lipotoxicity in HepG2 cells triggered by free fatty acids","volume":"3","author":"Yao","year":"2011","journal-title":"Am. J. Transl. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"42","DOI":"10.3389\/fnut.2017.00042","article-title":"The Nutraceutic Silybin Counteracts Excess Lipid Accumulation and Ongoing Oxidative Stress in an In Vitro Model of Non-Alcoholic Fatty Liver Disease Progression","volume":"4","author":"Vecchione","year":"2017","journal-title":"Front. Nutr."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.dib.2017.09.019","article-title":"Data on chow, liver tissue and mitochondrial fatty acid compositions as well as mitochondrial proteome changes after feeding mice a western diet for 6\u201324 weeks","volume":"15","author":"Einer","year":"2017","journal-title":"Data Br."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1089\/109793301753407948","article-title":"Nile red binding to HepG2 cells: An improved assay for in vitro studies of hepa-tosteatosis","volume":"14","author":"McMillian","year":"2001","journal-title":"Vitr. Mol. Toxicol. J. Basic Appl. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1112","DOI":"10.1038\/nprot.2006.179","article-title":"Sulforhodamine B colorimetric assay for cytotoxicity screening","volume":"1","author":"Vichai","year":"2006","journal-title":"Nat. Protoc."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1139\/y59-099","article-title":"A rapid method of total lipid extraction and purification","volume":"37","author":"Bligh","year":"1959","journal-title":"Can. J. Biochem. Physiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1007\/BF02531316","article-title":"Two dimensional thin layer chromatographic separation of polar lipids and determi-nation of phospholipids by phosphorus analysis of spots","volume":"5","author":"Rouser","year":"1970","journal-title":"Lipids"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"13843","DOI":"10.1074\/jbc.M808515200","article-title":"Loss of PINK1 function promotes mitophagy through effects on oxidative stress and mitochondrial fission","volume":"284","author":"Dagda","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1016\/S1046-2023(02)00038-5","article-title":"Blue Native electrophoresis to study mitochondrial and other protein complexes","volume":"26","author":"Nijtmans","year":"2002","journal-title":"Methods"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1002\/cptx.1","article-title":"Determination of metabolic viability and cell mass using a tandem resaz-urin\/sulforhodamine B assay","volume":"68","author":"Silva","year":"2016","journal-title":"Curr. Protoc. Toxicol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Ross, B.C. (2014). Mutual Information between Discrete and Continuous Data Sets. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0087357"},{"key":"ref_38","unstructured":"MacQueen, J. (July, January 21). Some methods for classification and analysis of multivariate observations. Proceedings of the Fifth Berkeley Symposium on Mathematical Statistics and Probability, Berkeley, CA, USA."},{"key":"ref_39","unstructured":"McKinney, W. (July, January 28). Data Structures for Statistical Computing in Python. Proceedings of the 9th Python in Science Conference, Austin, TX, USA."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1109\/MCSE.2011.37","article-title":"The NumPy array: A structure for efficient numerical computation","volume":"13","author":"Colbert","year":"2011","journal-title":"Comput. Sci. Eng."},{"key":"ref_41","unstructured":"Jones, E., Oliphant, T., and Peterson, P. (2021, April 12). SciPy: Open Source Scientific Tools for Python. Available online: http:\/\/www.scipy.org\/."},{"key":"ref_42","first-page":"2825","article-title":"Scikit-learn: Machine Learning in Python","volume":"12","author":"Pedregosa","year":"2011","journal-title":"J. Mach. Learn. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1016\/j.jhep.2018.01.019","article-title":"Fructose and sugar: A major mediator of non-alcoholic fatty liver disease","volume":"68","author":"Jensen","year":"2018","journal-title":"J. Hepatol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"21873","DOI":"10.3390\/ijms141121873","article-title":"Hepatic Adverse Effects of Fructose Consumption Independent of Overweight\/Obesity","volume":"14","author":"Schultz","year":"2013","journal-title":"Int. J. Mol. Sci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"G205","DOI":"10.1152\/ajpgi.00270.2015","article-title":"Defect of mitochondrial respiratory chain is a mechanism of ROS overproduction in a rat model of alcoholic liver disease: Role of zinc deficiency","volume":"310","author":"Sun","year":"2016","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1002\/hep.1840380426","article-title":"Defective hepatic mitochondrial respiratory chain in patients with nonal-coholic steatohepatitis","volume":"38","author":"Rubio","year":"2003","journal-title":"Hepatology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.cbi.2006.11.004","article-title":"A human hepatocellular in vitro model to investigate steatosis","volume":"165","author":"Donato","year":"2007","journal-title":"Chem. Biol. Interact."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1002\/mnfr.201500635","article-title":"Fructose and glucose combined with free fatty acids induce metabolic disorders in HepG2 cell: A new model to study the impacts of high-fructose\/sucrose and high-fat diets in vitro","volume":"60","author":"Zhao","year":"2016","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"104952","DOI":"10.1016\/j.tiv.2020.104952","article-title":"Fructose-palmitate based high calorie induce steatosis in HepG2 cells via mitochondrial dysfunction: An in vitro approach","volume":"68","author":"Sasi","year":"2020","journal-title":"Toxicol In Vitro"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"22474","DOI":"10.1016\/S0021-9258(18)45729-8","article-title":"Differential Regulation of Hepatic Triglyceride Lipase and Reductase Gene Expression in a Human Hepatoma Cell Line, HepG2","volume":"265","author":"Busch","year":"1990","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2504","DOI":"10.1194\/jlr.M039669","article-title":"Regulation of lipid droplet size and phospholipid composition by stearoyl-CoA desaturase","volume":"54","author":"Shi","year":"2013","journal-title":"J. Lipid Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1172\/JCI96702","article-title":"Fructose metabolism and metabolic disease","volume":"128","author":"Hannou","year":"2018","journal-title":"J. Clin. Investig."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1083\/jcb.201606069","article-title":"Mitochondrial lipid transport and biosynthesis: A complex balance","volume":"214","author":"Mesmin","year":"2016","journal-title":"J. Cell Biol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1111\/j.1432-1033.1978.tb12205.x","article-title":"Human liver mitochondria: Relation of a particular lipid composition to the mobility of spin-labelled lipids","volume":"84","author":"Benga","year":"1978","journal-title":"Eur. J. Biochem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1083\/jcb.201006159","article-title":"Making heads or tails of phospholipids in mitochondria","volume":"192","author":"Osman","year":"2011","journal-title":"J. Cell Biol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1016\/j.bbamcr.2010.09.014","article-title":"Apoptosis-induced changes in mitochondrial lipids","volume":"1813","author":"Crimi","year":"2011","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.cmet.2006.03.007","article-title":"The ratio of phosphatidylcholine to phosphatidylethanolamine influences membrane integrity and steatohepatitis","volume":"3","author":"Li","year":"2006","journal-title":"Cell Metab."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"2620","DOI":"10.2337\/db13-0993","article-title":"The Concentration of Phosphatidylethanolamine in Mitochondria Can Modulate ATP Production and Glucose Metabolism in Mice","volume":"63","author":"Lingrell","year":"2014","journal-title":"Diabetes"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1139\/apnm-2012-0261","article-title":"Nonalcoholic fatty liver disease is associated with lower hepatic and erythrocyte ratios of phosphatidylcholine to phosphatidylethanolamine","volume":"38","author":"Arendt","year":"2013","journal-title":"Appl. Physiol. Nutr. Metab."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1242\/jcs.071233","article-title":"Lipid map of the mammalian cell","volume":"124","year":"2011","journal-title":"J. Cell Sci."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"4158","DOI":"10.1074\/jbc.M112.434183","article-title":"Phosphatidylethanolamine Deficiency in Mammalian Mitochondria Impairs Oxida-tive Phosphorylation and Alters Mitochondrial Morphology","volume":"288","author":"Tasseva","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1038\/nature09968","article-title":"Aberrant lipid metabolism disrupts calcium homeostasis causing liver endoplasmic reticulum stress in obesity","volume":"473","author":"Fu","year":"2011","journal-title":"Nature"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1007\/s00125-015-3566-z","article-title":"Beta cell response to nutrient overload involves phospholipid remodelling and lipid peroxidation","volume":"58","author":"Cohen","year":"2015","journal-title":"Diabetologia"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1905","DOI":"10.1002\/hep.22239","article-title":"Prevention of free fatty acid-induced hepatic lipotoxicity by 18\u03b2-glycyrrhetinic acid through lysosomal and mitochondrial pathways","volume":"47","author":"Wu","year":"2008","journal-title":"Hepatology"},{"key":"ref_65","first-page":"198","article-title":"Dynamics of mitochondria in living cells","volume":"219","year":"1994","journal-title":"Microsc. Res. Tech."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.tem.2015.12.001","article-title":"Mitochondrial Dynamics and Metabolic Regulation","volume":"27","author":"Wai","year":"2016","journal-title":"Trends Endocrinol. Metab."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1089\/ars.2012.4779","article-title":"Mitochondrial Morphology in Metabolic Diseases","volume":"19","author":"Galloway","year":"2013","journal-title":"Antioxid. Redox Signal."},{"key":"ref_68","first-page":"183","article-title":"In vitro treatment of HepG2 cells with saturated fatty acids reproduces mitochondrial dysfunction found in nonalcoholic steatohepatitis","volume":"8","year":"2015","journal-title":"DMM Dis. Model Mech."},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Su\u00e1rez-Rivero, J.M., Villanueva-Paz, M., De La Cruz-Ojeda, P., De La Mata, M., Cot\u00e1n, D., Oropesa-\u00c1vila, M., De Lavera, I., \u00c1lvarez-C\u00f3rdoba, M., Luz\u00f3n-Hidalgo, R., and S\u00e1nchez-Alc\u00e1zar, J.A. (2016). Mitochondrial Dynamics in Mitochondrial Diseases. Diseases, 5.","DOI":"10.3390\/diseases5010001"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1476-511X-11-142","article-title":"Conjugated linoleic acid or omega 3 fatty acids increase mitochondrial bio-synthesis and metabolism in skeletal muscle cells","volume":"11","author":"Vaughan","year":"2012","journal-title":"Lipids Health Dis."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"317","DOI":"10.3746\/pnf.2016.21.4.317","article-title":"Effects of Eicosapentaenoic Acid and Docosahexaenoic Acid on Mitochondrial DNA Replication and PGC-1\u03b1 Gene Expression in C2C12Muscle Cells","volume":"21","author":"Lee","year":"2016","journal-title":"Prev. Nutr. Food Sci."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"1439","DOI":"10.1159\/000358709","article-title":"Palmitate-Induced Cell Death and Mitochondrial Respiratory Dysfunction in Myoblasts are Not Prevented by Mitochondria-Targeted Antioxidants","volume":"33","author":"Trnka","year":"2014","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1111\/j.1582-4934.2010.01128.x","article-title":"High fat diet-induced liver steatosis promotes an increase in liver mitochondrial biogenesis in response to hypoxia","volume":"15","author":"Carabelli","year":"2010","journal-title":"J. Cell. Mol. Med."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.mito.2012.10.011","article-title":"Is mitochondrial DNA content a potential biomarker of mitochondrial dysfunction?","volume":"13","author":"Malik","year":"2013","journal-title":"Mitochondrion"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1007\/s10863-012-9448-x","article-title":"Dietary fatty acids modulate liver mitochondrial cardiolipin content and its fatty acid composition in rats with non alcoholic fatty liver disease","volume":"44","author":"Aoun","year":"2012","journal-title":"J. Bioenerg. Biomembr."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"368","DOI":"10.1017\/S0007114517003713","article-title":"20-Week follow-up of hepatic steatosis installation and liver mitochondrial structure and activity and their interrelation in rats fed a high-fat\u2013high-fructose diet","volume":"119","author":"Fouret","year":"2018","journal-title":"Br. J. Nutr."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/S0014-5793(01)03206-9","article-title":"Reactive oxygen species generated from the mitochondrial electron transport chain induce cytochrome c dissociation from beef-heart submitochondrial particles via cardiolipin peroxidation. Possible role in the apoptosis","volume":"509","author":"Petrosillo","year":"2001","journal-title":"FEBS Lett."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1007\/s11306-014-0729-8","article-title":"Temporal metabolomic responses of cultured HepG2 liver cells to high fructose and high glucose exposures","volume":"11","author":"Meissen","year":"2015","journal-title":"Metabolomics"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"e291","DOI":"10.1038\/emm.2016.157","article-title":"Oxidative stress and calcium dysregulation by palmitate in type 2 diabetes","volume":"49","author":"Ly","year":"2017","journal-title":"Exp. Mol. Med."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1203\/00006450-199209000-00024","article-title":"The Effect of Monosaturated and Polyunsaturated Fatty Acids on Oxygen Toxicity in Cultured Cells","volume":"32","author":"Spitz","year":"1992","journal-title":"Pediatr. Res."},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Joseph, L.C., Barca, E., Subramanyam, P., Komrowski, M., Pajvani, U., Colecraft, H.M., Hirano, M., and Morrow, J.P. (2016). Inhibition of NAPDH Oxidase 2 (NOX2) Prevents Oxidative Stress and Mitochondrial Abnormalities Caused by Saturated Fat in Cardiomyocytes. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0145750"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"16871","DOI":"10.1073\/pnas.0809255105","article-title":"Differential effects of central fructose and glucose on hypothalamic malonyl\u2014CoA and food intake","volume":"105","author":"Cha","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1038\/s41575-018-0065-y","article-title":"Apoptosis and necroptosis in the liver: A matter of life and death","volume":"15","author":"Schwabe","year":"2018","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1080\/01926230701320337","article-title":"Apoptosis: A review of programmed cell death","volume":"35","author":"Elmore","year":"2007","journal-title":"Toxicol. Pathol."},{"key":"ref_85","doi-asserted-by":"crossref","unstructured":"Brentnall, M., Rodriguez-Menocal, L., De Guevara, R.L., Cepero, E., and Boise, L.H. (2013). Caspase-9, caspase-3 and caspase-7 have distinct roles during intrinsic apoptosis. BMC Cell Biol., 14.","DOI":"10.1186\/1471-2121-14-32"},{"key":"ref_86","first-page":"1825","article-title":"Biochimica et Biophysica Acta The molecular mechanism of apoptosis upon caspase-8 activation: Quantitative experimental validation of a mathematical model","volume":"1823","author":"Kominami","year":"2012","journal-title":"BBA Mol. Cell Res."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1194\/jlr.R066357","article-title":"Lipotoxic lethal and sublethal stress signaling in hepatocytes: Relevance to NASH pathogenesis","volume":"57","author":"Hirsova","year":"2016","journal-title":"J. Lipid Res."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1038\/sj.cdd.4401457","article-title":"Cardiolipin and its metabolites move from mitochondria to other cellular membranes during death receptor-mediated apoptosis","volume":"11","author":"Sorice","year":"2004","journal-title":"Cell Death Differ."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1042\/BJ20041389","article-title":"Pro-apoptotic Bid induces membrane perturbation by inserting selected lysolipids into the bilayer","volume":"387","author":"Goonesinghe","year":"2005","journal-title":"Biochem. J."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"2199","DOI":"10.1016\/j.bpj.2010.01.037","article-title":"Understanding detergent effects on lipid membranes: A model study of lysolipids","volume":"98","author":"Henriksen","year":"2010","journal-title":"Biophys. J."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/S0092-8674(03)00422-7","article-title":"Apoptotic Cells Induce Migration of Phagocytes via Caspase-3-Mediated Release of a Lipid Attraction Signal","volume":"113","author":"Lauber","year":"2003","journal-title":"Cell"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"2331","DOI":"10.2337\/dc12-1760","article-title":"Circulating Lysophosphatidylcholines Are Markers of a Metabolically Benign Nonalcoholic Fatty Liver","volume":"36","author":"Lehmann","year":"2013","journal-title":"Diabetes Care"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"81","DOI":"10.3389\/fimmu.2018.00081","article-title":"Adipocyte Fatty Acid-Binding Protein Promotes Palmitate-Induced Mitochondrial Dysfunction and Apoptosis in Macrophages","volume":"9","author":"Li","year":"2018","journal-title":"Front. Immunol."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"a026716","DOI":"10.1101\/cshperspect.a026716","article-title":"Caspase Functions in Cell Death and Disease: Figure 1","volume":"7","author":"McIlwain","year":"2015","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_95","first-page":"S637","article-title":"The Effect of D-Galactosamine on Lean and Steatotic Rat Hepatocytes in Primary Culture","volume":"64","author":"Lotkova","year":"2015","journal-title":"Physiol. Res."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/j.lfs.2018.04.022","article-title":"Oleic acid protects saturated fatty acid mediated lipotoxicity in hepatocytes and rat of non-alcoholic steatohepatitis","volume":"203","author":"Chen","year":"2018","journal-title":"Life Sci."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.1002\/hep.22737","article-title":"Unsaturated fatty acids inhibit the expression of tumor suppressor phosphatase and tensin homolog (PTEN) via microRNA-21 up-regulation in hepatocytes","volume":"49","author":"Vinciguerra","year":"2008","journal-title":"Hepatology"}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/5\/1723\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:04:11Z","timestamp":1760162651000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/13\/5\/1723"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,19]]},"references-count":97,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["nu13051723"],"URL":"https:\/\/doi.org\/10.3390\/nu13051723","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,19]]}}}