Dedifferentiation of Primary Hepatocytes is Accompanied with Reorganization of Lipid Metabolism Indicated by Altered Molecular Lipid and miRNA Profiles
Abstract
:1. Introduction
2. Results
2.1. The Morphology, Gene Expression, and Functionality of PHHs
2.2. Alterations in the Lipid Profile of the PHHs
2.3. Alterations in the Lipid Metabolism-Related Genes
2.4. Alterations in FA Metabolism-Related Genes
2.5. Alterations in the Genes Related to Glucose Homeostasis
2.6. Alterations in the miRNA Profile of the PHHs
3. Discussion
4. Method and Materials
4.1. Cell Culture
4.2. Real-Time Quantitative PCR Analysis
4.3. Triacylglycerol, Albumin, and Urea Secretion
4.4. Lipid Mass Spectrometry
4.5. miRNA Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
CE | cholesteryl ester |
CerS | ceramide synthase |
DAG | diacylglycerol |
ELOVL | fatty acid elongase |
ER | endoplasmic reticulum |
FA | fatty acid |
FADS | fatty acid desaturase |
FASN | fatty acid synthase |
Gb3 | globotriaosylceramide |
GCK | glucokiase |
Glc/GalCer | glucosyl/galactosylceramide |
GSL | glycosphingolipid |
hLTR | human liver total RNA |
HUFA | highly unsaturated fatty acid |
LacCer | lactosylceramide |
LCFA | long-chain fatty acid |
miRNA | microRNA |
MUFA | monounsaturated fatty acid |
PC | phosphatidylcholine |
PE | phosphatidylethanolamine |
PCK1 | phosphoenolpyruvate carboxykinase |
PHH | primary human hepatocyte |
PI | phosphatidylinositol |
PKLR | liver-type pyruvate kinase |
PL | phospholipid |
PS | phosphatidylserine |
PUFA | polyunsaturated fatty acid |
qPCR | quantitative PCR |
RXRα | retinoid X receptor alpha |
SCD | stearoyl-CoA desaturase |
SFA | saturated fatty acid |
SL | sphingolipid |
SM | sphingomyelin |
SMS | sphingomyelin synthase |
TAG | triacylglycerol |
UGCG | UDP-glucose ceramide glucosyltransferase |
VLCFA | very-long-chain fatty acid |
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Genes | Functions | References |
---|---|---|
CERS1, CERS2, CERS4, CERS5, CERS6 | Encoding enzymes that catalyze the synthesis of ceramides from sphingoid base and acyl-CoA substrates. | [33,44,45] |
ASAH1, ASAH2 | Encoding acid (ASAH1) and neutral (ASAH2) ceramidases that degrade ceramides to sphingosine and free fatty acids (FAs). | [29] |
SGMS1, SGMS2 | Synthesis of sphingomyelin from ceramides and crucial in ceramide and sphingomyelin homoeostasis. | [46] |
SMPD1 | Encoding a lysosomal acid sphingomyelinase hydrolyzing sphingomyelin to ceramide. | [47] |
UGCG | Encoding the enzyme that catalyzes the first glycosylation step in glycosphingolipid biosynthesis. | [48] |
FADS1, FADS2 | Encoding for the enzymes inserting double bonds to pre-existing polyunsaturated fatty acid (PUFA) precursors. Critical for the synthesis and regulation of PUFAs. | [49] |
ELOVL1, ELOVL2, ELOVL3, ELOVL5, ELOVL6, ELOVL7 | Encoding enzymes that function in concert with FADSs and determine the rate of overall fatty acid elongation. ELOVL1, ELOVL3, ELOVL6, and ELOVL7 are involved in elongation of saturated and monounsaturated FAs, while ELOVL2 and ELOVL5 are involved in elongation of PUFAs. ELOVL5 also elongates some monounsaturated FAs, such as palmitoleic acid (FA 16:1). | [50] |
FASN | Encoding the enzyme that determines the de-novo synthesis of FAs by synthesising palmitic acid (FA 16:0). | [40] |
SCD | Encoding a rate-limiting enzyme in the cellular synthesis of monounsaturated FAs from saturated FAs. | [43] |
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Kiamehr, M.; Heiskanen, L.; Laufer, T.; Düsterloh, A.; Kahraman, M.; Käkelä, R.; Laaksonen, R.; Aalto-Setälä, K. Dedifferentiation of Primary Hepatocytes is Accompanied with Reorganization of Lipid Metabolism Indicated by Altered Molecular Lipid and miRNA Profiles. Int. J. Mol. Sci. 2019, 20, 2910. https://doi.org/10.3390/ijms20122910
Kiamehr M, Heiskanen L, Laufer T, Düsterloh A, Kahraman M, Käkelä R, Laaksonen R, Aalto-Setälä K. Dedifferentiation of Primary Hepatocytes is Accompanied with Reorganization of Lipid Metabolism Indicated by Altered Molecular Lipid and miRNA Profiles. International Journal of Molecular Sciences. 2019; 20(12):2910. https://doi.org/10.3390/ijms20122910
Chicago/Turabian StyleKiamehr, Mostafa, Laura Heiskanen, Thomas Laufer, Aneta Düsterloh, Mustafa Kahraman, Reijo Käkelä, Reijo Laaksonen, and Katriina Aalto-Setälä. 2019. "Dedifferentiation of Primary Hepatocytes is Accompanied with Reorganization of Lipid Metabolism Indicated by Altered Molecular Lipid and miRNA Profiles" International Journal of Molecular Sciences 20, no. 12: 2910. https://doi.org/10.3390/ijms20122910