Please use this identifier to cite or link to this item: https://hdl.handle.net/10316/105302
Title: Exploratory Data Analysis of Cell and Mitochondrial High-Fat, High-Sugar Toxicity on Human HepG2 Cells
Authors: Amorim, Ricardo 
Simões, Inês C. M.
Veloso, Caroline 
Carvalho, Adriana 
Simões, Rui F. 
Pereira, Francisco B. 
Thiel, Theresa
Normann, Andrea
Morais, Catarina M. 
Jurado, Maria Amália da Silva 
Wieckowski, Mariusz R. 
Teixeira, José 
Oliveira, Paulo J. 
Keywords: non-alcoholic fatty liver disease (NAFLD); in vitro cell model; Hepg2 cells; lipid accumulation; mitochondria dys(function); exploratory data analysis
Issue Date: 19-May-2021
Publisher: MDPI
Project: PTDC/BTM-SAL/29297/2017 
POCI-01-0145-FEDER-029297 
PTDC/DTPFTO/ 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-029152 
UIDB/04539/2020 
SFRH/BD/131070/2017 
NORTE-01-0145-FEDER-00002 
National Science Centre, Poland (UMO-2018/29/B/NZ1/00589) 
Marie Skłodowska-Curie Grant Agreement No. 722619 (FOIE GRAS) and Grant Agreement No. 734719 (mtFOIE GRAS) 
Serial title, monograph or event: Nutrients
Volume: 13
Issue: 5
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.
URI: https://hdl.handle.net/10316/105302
ISSN: 2072-6643
DOI: 10.3390/nu13051723
Rights: openAccess
Appears in Collections:I&D CNC - Artigos em Revistas Internacionais
IIIUC - Artigos em Revistas Internacionais
I&D CISUC - Artigos em Revistas Internacionais

Show full item record

SCOPUSTM   
Citations

7
checked on May 6, 2024

WEB OF SCIENCETM
Citations

7
checked on May 2, 2024

Page view(s)

59
checked on May 21, 2024

Download(s)

14
checked on May 21, 2024

Google ScholarTM

Check

Altmetric

Altmetric


This item is licensed under a Creative Commons License Creative Commons