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An effective protocol for pharmacological defatting of primary human hepatocytes which is non-toxic to cholangiocytes or intrahepatic endothelial cells.
- Source :
-
PloS one [PLoS One] 2018 Jul 25; Vol. 13 (7), pp. e0201419. Date of Electronic Publication: 2018 Jul 25 (Print Publication: 2018). - Publication Year :
- 2018
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Abstract
- Introduction: Pharmacological defatting of rat hepatocytes and hepatoma cell lines suggests that the same method could be used to ameliorate macrovesicular steatosis in moderate to severely fatty livers. However there is no data assessing the effects of those drugs on primary human liver cells. We aimed to determine the effectiveness of a pharmacological cocktail in reducing the in vitro lipid content of primary human hepatocytes (PHH). In addition we sought to determine the cytotoxicity of the cocktail towards non-parenchymal liver cells.<br />Methods: Steatosis was induced in PHH by supplementation with a combination of saturated and unsaturated free fatty acids. This was followed by addition of a defatting drug cocktail for up to 48 hours. The same experimental method was used with human intra-hepatic endothelial cells (HIEC) and human cholangiocytes. MTT assay was used to assess cell viability, triglyceride quantification and oil red O staining were used to determine intracellular lipids content whilst ketone bodies were measured in the supernatants following experimentation.<br />Results: Incubation of fat loaded PHH with the drugs over 48 hours reduced the intracellular lipid area by 54%, from 12.85% to 5.99% (p = 0.002) (percentage of total oil red O area), and intracellular triglyceride by 35%, from 28.24 to 18.30 nmol/million of cells (p<0.001). Total supernatant ketone bodies increased 1.4-fold over 48 hours in the defatted PHH compared with vehicle controls (p = 0.002). Moreover incubation with the drugs for 48 hours increased the viability of PHH by 11%, cholangiocytes by 25% whilst having no cytotoxic effects on HIEC.<br />Conclusion: These data demonstrate that pharmacological intervention can significantly decrease intracellular lipid content of PHH, increase fatty acids β-oxidation whilst being non-toxic to PHH, HIEC or cholangiocytes.<br />Competing Interests: The authors have declared that no competing interests exist.
- Subjects :
- Bile Ducts cytology
Bile Ducts drug effects
Bile Ducts metabolism
Cell Survival drug effects
Cells, Cultured
Endothelial Cells cytology
Endothelial Cells drug effects
Endothelial Cells metabolism
Epithelial Cells cytology
Epithelial Cells drug effects
Epithelial Cells metabolism
Fatty Acids metabolism
Fatty Liver metabolism
Hep G2 Cells
Hepatocytes cytology
Hepatocytes metabolism
Humans
Ketone Bodies metabolism
Liver cytology
Liver metabolism
Fats metabolism
Fatty Liver drug therapy
Hepatocytes drug effects
Liver drug effects
Triglycerides metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 13
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 30044872
- Full Text :
- https://doi.org/10.1371/journal.pone.0201419