Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells

Ricardo Amorim; Inês Simões; Caroline Veloso; Adriana Carvalho; Rui Simões; Francisco Pereira; Theresa Thiel; Andrea Normann; Catarina Morais; Amália Jurado; Mariusz Wieckowski; José Teixeira; Paulo Oliveira

doi:10.3390/nu13051723

journal article Open Access May 19, 2021

Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells

Ricardo Amorim Inês Simões

Caroline Veloso

Adriana Carvalho Rui Simões Francisco Pereira Theresa Thiel Andrea Normann Catarina Morais Amália Jurado Mariusz Wieckowski José Teixeira

Paulo Oliveira

Nutrients Vol. 13 No. 5 pp. 1723 · MDPI AG

View at Publisher Save 10.3390/nu13051723

Abstract

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.

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Details

Published: May 19, 2021
Vol/Issue: 13(5)
Pages: 1723
License: View

Authors

R

Ricardo Amorim

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; CIQUP/Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal; PhD Programme in Experimental Biology and Biomedicine (PDBEB), Institute for Interdisciplinary Research (IIIUC), University of Coimbra, 3004-531 Coimbra, Portugal

I

Inês Simões

Laboratory of Mitochondrial Biology and Metabolism, Nencki Institute of Experimental Biology of Polish Academy of Sciences, 02-093 Warsaw, Poland

C

Caroline Veloso

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

A

Adriana Carvalho

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; PhD Programme in Experimental Biology and Biomedicine (PDBEB), Institute for Interdisciplinary Research (IIIUC), University of Coimbra, 3004-531 Coimbra, Portugal

R

Rui Simões

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; PhD Programme in Experimental Biology and Biomedicine (PDBEB), Institute for Interdisciplinary Research (IIIUC), University of Coimbra, 3004-531 Coimbra, Portugal

F

Francisco Pereira

Center for Informatics and Systems, University of Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra, Portugal; Coimbra Polytechnic-ISEC, 3030-190 Coimbra, Portugal

T

Theresa Thiel

Mediagnostic, D-72770 Reutlingen, Germany

A

Andrea Normann

Mediagnostic, D-72770 Reutlingen, Germany

C

Catarina Morais

Center for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal

A

Amália Jurado

Center for Neuroscience and Cell Biology, Department of Life Sciences, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal

M

Mariusz Wieckowski

Laboratory of Mitochondrial Biology and Metabolism, Nencki Institute of Experimental Biology of Polish Academy of Sciences, 02-093 Warsaw, Poland

J

José Teixeira

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; CIQUP/Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto, 4169-007 Porto, Portugal

P

Paulo Oliveira

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

Funding

H2020 Marie Skłodowska-Curie Actions Award: 722619, 734719

Narodowe Centrum Nauki Award: UMO-2018/29/B/NZ1/00589

Fundação para a Ciência 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

Cite This Article

Ricardo Amorim, Inês Simões, Caroline Veloso, et al. (2021). Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells. Nutrients, 13(5), 1723. https://doi.org/10.3390/nu13051723

Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells

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