Journal article
Modifying nutritional substrates induces macrovesicular lipid droplet accumulation and metabolic alterations in a cellular model of hepatic steatosis
- Abstract:
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Background and Aims: Nonalcoholic fatty liver disease (NAFLD) begins with steatosis, where a mixed macrovesicular pattern of large and small lipid droplets (LDs) develops. Since in vitro models recapitulating this are limited, the aims of this study were to develop mixed macrovesicular steatosis in immortalized hepatocytes and investigate effects on intracellular metabolism by altering nutritional substrates.
Methods: Huh7 cells were cultured in 11 mM glucose and 2% human serum (HS) for 7 days before additional sugars and fatty acids (FAs), either with 200 µM FAs (low fat low sugar; LFLS), 5.5 mM fructose + 200 µM FAs (low fat high sugar; LFHS), or 5.5 mM fructose + 800 µM FAs (high fat high sugar; HFHS), were added for 7 days. FA metabolism, lipid droplet characteristics, and transcriptomic signatures were investigated.
Results: Between the LFLS and LFHS conditions, there were few notable differences. In the HFHS condition, intracellular triacylglycerol (TAG) was increased and the LD pattern and distribution was similar to that found in primary steatotic hepatocytes. HFHS‐treated cells had lower levels of de novo‐derived FAs and secreted larger, TAG‐rich lipoprotein particles. RNA sequencing and gene set enrichment analysis showed changes in several pathways including those involved in metabolism and cell cycle.
Conclusions: Repeated doses of HFHS treatment resulted in a cellular model of NAFLD with a mixed macrovesicular LD pattern and metabolic dysfunction. Since these nutrients have been implicated in the development of NAFLD in humans, the model provides a good physiological basis for studying NAFLD development or regression in vitro.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, 2.0MB, Terms of use)
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- Publisher copy:
- 10.14814/phy2.14482
Authors
- Publisher:
- Wiley Open Access
- Journal:
- Physiological Reports More from this journal
- Volume:
- 8
- Issue:
- 13
- Article number:
- e14482
- Publication date:
- 2020-07-08
- Acceptance date:
- 2020-05-18
- DOI:
- ISSN:
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2051-817X
- Language:
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English
- Keywords:
- Pubs id:
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1105506
- Local pid:
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pubs:1105506
- Deposit date:
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2020-05-19
Terms of use
- Copyright holder:
- Gunn et al.
- Copyright date:
- 2020
- Rights statement:
- © 2020 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Licence:
- CC Attribution (CC BY)
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