Comparison of HepaRG and HepG2 cell lines to model mitochondrial respiratory adaptations in non‑alcoholic fatty liver disease

Maseko,Tumisang Edward; Elkalaf,Moustafa; Peterová,Eva; Lotková,Halka; Staňková,Pavla; Melek,Jan; Dušek,Jan; Žádníková,Petra; Čížková,Dana; Bezrouk,Aleš; Pávek,Petr; Červinková,Zuzana; Kučera,Otto

doi:10.3892/ijmm.2023.5342

Comparison of HepaRG and HepG2 cell lines to model mitochondrial respiratory adaptations in non‑alcoholic fatty liver disease

Authors:
- Tumisang Edward Maseko
- Moustafa Elkalaf
- Eva Peterová
- Halka Lotková
- Pavla Staňková
- Jan Melek
- Jan Dušek
- Petra Žádníková
- Dana Čížková
- Aleš Bezrouk
- Petr Pávek
- Zuzana Červinková
- Otto Kučera
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Affiliations: Department of Physiology, Charles University, Faculty of Medicine in Hradec Kralove, 500 03 Hradec Kralove, Czech Republic, Department of Histology and Embryology, Charles University, Faculty of Medicine in Hradec Kralove, 500 03 Hradec Kralove, Czech Republic, Department of Medical Biophysics, Charles University, Faculty of Medicine in Hradec Kralove, 500 03 Hradec Kralove, Czech Republic, Department of Pharmacology and Toxicology, Charles University, Faculty of Pharmacy in Hradec Kralove, 500 05 Hradec Kralove, Czech Republic
Published online on: December 22, 2023 https://doi.org/10.3892/ijmm.2023.5342
Article Number: 18
Copyright: © Maseko et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Although some clinical studies have reported increased mitochondrial respiration in patients with fatty liver and early non‑alcoholic steatohepatitis (NASH), there is a lack of in vitro models of non‑alcoholic fatty liver disease (NAFLD) with similar findings. Despite being the most commonly used immortalized cell line for in vitro models of NAFLD, HepG2 cells exposed to free fatty acids (FFAs) exhibit a decreased mitochondrial respiration. On the other hand, the use of HepaRG cells to study mitochondrial respiratory changes following exposure to FFAs has not yet been fully explored. Therefore, the present study aimed to assess cellular energy metabolism, particularly mitochondrial respiration, and lipotoxicity in FFA‑treated HepaRG and HepG2 cells. HepaRG and HepG2 cells were exposed to FFAs, followed by comparative analyses that examained cellular metabolism, mitochondrial respiratory enzyme activities, mitochondrial morphology, lipotoxicity, the mRNA expression of selected genes and triacylglycerol (TAG) accumulation. FFAs stimulated mitochondrial respiration and glycolysis in HepaRG cells, but not in HepG2 cells. Stimulated complex I, II‑driven respiration and β‑oxidation were linked to increased complex I and II activities in FFA‑treated HepaRG cells, but not in FFA‑treated HepG2 cells. Exposure to FFAs disrupted mitochondrial morphology in both HepaRG and HepG2 cells. Lipotoxicity was induced to a greater extent in FFA‑treated HepaRG cells than in FFA‑treated HepG2 cells. TAG accumulation was less prominent in HepaRG cells than in HepG2 cells. On the whole, the present study demonstrates that stimulated mitochondrial respiration is associated with lipotoxicity in FFA‑treated HepaRG cells, but not in FFA‑treated HepG2 cells. These findings suggest that HepaRG cells are more suitable for assessing mitochondrial respiratory adaptations in the developed in vitro model of early‑stage NASH.

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