Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells
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...
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MDPI AG
2021-05-01
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Online Access: | https://www.mdpi.com/2072-6643/13/5/1723 |
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doaj-59bddf1a69cf40e3aeda99b0af4bca57 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ricardo Amorim Inês C. M. Simões Caroline Veloso Adriana Carvalho Rui F. Simões Francisco B. Pereira Theresa Thiel Andrea Normann Catarina Morais Amália S. Jurado Mariusz R. Wieckowski José Teixeira Paulo J. Oliveira |
spellingShingle |
Ricardo Amorim Inês C. M. Simões Caroline Veloso Adriana Carvalho Rui F. Simões Francisco B. Pereira Theresa Thiel Andrea Normann Catarina Morais Amália S. Jurado Mariusz R. Wieckowski José Teixeira Paulo J. Oliveira Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells Nutrients non-alcoholic fatty liver disease (NAFLD) in vitro cell model Hepg2 cells lipid accumulation mitochondria dys(function) exploratory data analysis |
author_facet |
Ricardo Amorim Inês C. M. Simões Caroline Veloso Adriana Carvalho Rui F. Simões Francisco B. Pereira Theresa Thiel Andrea Normann Catarina Morais Amália S. Jurado Mariusz R. Wieckowski José Teixeira Paulo J. Oliveira |
author_sort |
Ricardo Amorim |
title |
Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells |
title_short |
Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells |
title_full |
Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells |
title_fullStr |
Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells |
title_full_unstemmed |
Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells |
title_sort |
exploratory data analysis of cell and mitochondrial high-fat, high-sugar toxicity on human hepg2 cells |
publisher |
MDPI AG |
series |
Nutrients |
issn |
2072-6643 |
publishDate |
2021-05-01 |
description |
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 (ΔΨm) 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. |
topic |
non-alcoholic fatty liver disease (NAFLD) in vitro cell model Hepg2 cells lipid accumulation mitochondria dys(function) exploratory data analysis |
url |
https://www.mdpi.com/2072-6643/13/5/1723 |
work_keys_str_mv |
AT ricardoamorim exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT inescmsimoes exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT carolineveloso exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT adrianacarvalho exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT ruifsimoes exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT franciscobpereira exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT theresathiel exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT andreanormann exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT catarinamorais exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT amaliasjurado exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT mariuszrwieckowski exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT joseteixeira exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells AT paulojoliveira exploratorydataanalysisofcellandmitochondrialhighfathighsugartoxicityonhumanhepg2cells |
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doaj-59bddf1a69cf40e3aeda99b0af4bca572021-06-01T00:30:25ZengMDPI AGNutrients2072-66432021-05-01131723172310.3390/nu13051723Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 CellsRicardo Amorim0Inês C. M. Simões1Caroline Veloso2Adriana Carvalho3Rui F. Simões4Francisco B. Pereira5Theresa Thiel6Andrea Normann7Catarina Morais8Amália S. Jurado9Mariusz R. Wieckowski10José Teixeira11Paulo J. Oliveira12CNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, PortugalLaboratory of Mitochondrial Biology and Metabolism, Nencki Institute of Experimental Biology of Polish Academy of Sciences, 02-093 Warsaw, PolandCNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, PortugalCNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, PortugalCNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, PortugalCenter for Informatics and Systems, University of Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra, PortugalMediagnostic, D-72770 Reutlingen, GermanyMediagnostic, D-72770 Reutlingen, GermanyCenter for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, PortugalCenter for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, PortugalLaboratory of Mitochondrial Biology and Metabolism, Nencki Institute of Experimental Biology of Polish Academy of Sciences, 02-093 Warsaw, PolandCNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, PortugalCNC-Center for Neuroscience and Cell Biology, CIBB-Centre for Innovative Biomedicine and Biotechnology, University of Coimbra, UC-Biotech, Biocant Park, 3060-197 Cantanhede, PortugalNon-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 (ΔΨm) 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.https://www.mdpi.com/2072-6643/13/5/1723non-alcoholic fatty liver disease (NAFLD)in vitro cell modelHepg2 cellslipid accumulationmitochondria dys(function)exploratory data analysis |