Time-Dependent Changes in Hepatic Sphingolipid Accumulation and PI3K/Akt/mTOR Signaling Pathway in a Rat Model of NAFLD
Abstract
:1. Introduction
2. Results
2.1. Effects of Chronic High-Fat Chow Administration on the Hepatic Sphingolipid Concentration
2.2. Effects of Chronic High-Fat Chow Administration on the Hepatic Expression of Proteins from Sphingolipid Pathways
2.3. Effects of Chronic High-Fat Chow Administration on the Hepatic Expression or Phosphorylation State of Proteins Involved in the Insulin Signaling Pathway
3. Discussion
4. Materials and Methods
4.1. Experimental Model
4.2. Immunoblotting
4.3. Sphingolipids Analysis
4.4. Intracellular Content of Phosphoproteins
4.5. Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
Abbreviations
Alk-SMase | alkaline sphingomyelinase |
ASAH1 | acid ceramidase |
ASAH2 | neutral ceramidase |
ASAH3 | alkaline ceramidase |
BAD | Bcl-2-associated agonist of cell death |
BCA | bicinchoninic acid |
BSA | bovine serum albumin |
CER | ceramide |
CerS2, 4, 6 | ceramide synthase 2, 4, 6 |
FA | fatty acid |
FFA | free fatty acid |
GLC | gas–liquid chromatography |
GLUT | glucose transporter |
GSK3α/β | glycogen synthase 3 α/β |
HCC | hepatocellular carcinoma |
HFD | high-fat diet |
HPLC | high-performance liquid chromatography |
HRP | horseradish peroxidase |
IR | insulin resistance |
IRS | insulin receptor substrate |
mTOR | mechanistic target of rapamycin |
NAFLD | non-alcoholic fatty liver disease |
NASH | non-alcoholic steatohepatitis |
N-SMase | neutral sphingomyelinase |
P70 S6 Kinase | P70 ribosomal protein S6 kinase |
P90 S6 Kinase | P90 ribosomal protein S6 kinase |
PIP3 | phosphatidylinositol-3,4,5-triphosphate |
PI3K | phosphatidylinositol-3,4,5-triphosphate kinase |
PKB/Akt | protein kinase B |
PMSF | phenylmethylsulfonyl fluoride |
PTEN | phosphatase and tensin homolog |
PVDF | polyvinylidene fluoride |
S6RP | S6 ribosomal protein |
S1P | sphingosine-1-phosphate |
SA1P | sphinganine-1-phosphate |
SA-PE | streptavidin-phycoerythrin |
SD | standard deviation |
SDS-PAGE | sodium dodecyl sulfate-polyacrylamide gel electrophoresis |
SFA | sphinganine |
SFO | sphingosine |
SPHK1, 2 | sphingosine kinase 1, 2 |
SPTLC2 | serine palmitoyltransferase 2 |
TAG | triacylglycerol |
TBST | tris-buffered saline with Tween-20 |
References
- Park, W.-J.; Song, J.-H.; Kim, G.-T.; Park, T.-S. Ceramide and Sphingosine 1-Phosphate in Liver Diseases. Mol. Cells 2020, 43, 419–430. [Google Scholar]
- Choi, S.; Snider, A.J. Sphingolipids in High Fat Diet and Obesity-Related Diseases. Mediat. Inflamm. 2015, 2015, 520618. [Google Scholar] [CrossRef] [Green Version]
- Konstantynowicz-Nowicka, K.; Berk, K.; Chabowski, A.; Kasacka, I.; Bielawiec, P.; Łukaszuk, B.; Harasim-Symbor, E. High-Fat Feeding in Time-Dependent Manner Affects Metabolic Routes Leading to Nervonic Acid Synthesis in NAFLD. Int. J. Mol. Sci. 2019, 20, 3829. [Google Scholar] [CrossRef] [Green Version]
- Presa, N.; Clugston, R.D.; Lingrell, S.; Kelly, S.E.; Merrill, A.H.; Jana, S.; Kassiri, Z.; Gómez-Muñoz, A.; Vance, D.E.; Jacobs, R.L.; et al. Vitamin E alleviates non-alcoholic fatty liver disease in phosphatidylethanolamine N-methyltransferase deficient mice. Biochim. Biophys. Acta (BBA)—Mol. Basis Dis. 2019, 1865, 14–25. [Google Scholar] [CrossRef]
- Svegliati-Baroni, G.; Pierantonelli, I.; Torquato, P.; Marinelli, R.; Ferreri, C.; Chatgilialoglu, C.; Bartolini, D.; Galli, F. Lipidomic biomarkers and mechanisms of lipotoxicity in non-alcoholic fatty liver disease. Free Radic. Biol. Med. 2019, 144, 293–309. [Google Scholar] [CrossRef] [PubMed]
- Pei, K.; Gui, T.; Kan, D.; Feng, H.; Jin, Y.; Yang, Y.; Zhang, Q.; Du, Z.; Gai, Z.; Wu, J.; et al. An Overview of Lipid Metabolism and Nonalcoholic Fatty Liver Disease. BioMed Res. Int. 2020, 2020, 4020249. [Google Scholar] [CrossRef] [PubMed]
- Régnier, M.; Polizzi, A.; Guillou, H.; Loiseau, N. Sphingolipid metabolism in non-alcoholic fatty liver diseases. Biochimie 2019, 159, 9–22. [Google Scholar] [CrossRef] [PubMed]
- Gruben, N.; Shiri-Sverdlov, R.; Koonen, D.P.; Hofker, M.H. Nonalcoholic fatty liver disease: A main driver of insulin resistance or a dangerous liaison? Biochim. Biophys. Acta (BBA)—Mol. Basis Dis. 2014, 1842, 2329–2343. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Patterson, R.E.; Kalavalapalli, S.; Williams, C.M.; Nautiyal, M.; Mathew, J.T.; Martinez, J.; Reinhard, M.K.; McDougall, D.J.; Rocca, J.R.; Yost, R.A.; et al. Lipotoxicity in steatohepatitis occurs despite an increase in tricarboxylic acid cycle activity. Am. J. Physiol. Metab. 2016, 310, E484–E494. [Google Scholar] [CrossRef] [Green Version]
- Gai, Z.; Gui, T.; Alecu, I.; Lone, M.; Hornemann, T.; Chen, Q.; Visentin, M.; Hiller, C.; Hausler, S.; Kullak-Ublick, G.A. Farnesoid X receptor activation induces the degradation of hepatotoxic 1-deoxysphingolipids in non-alcoholic fatty liver disease. Liver Int. 2019, 40, 844–859. [Google Scholar] [CrossRef] [PubMed]
- Zabielski, P.; Hady, H.R.; Chacinska, M.; Roszczyc, K.; Górski, J.; Blachnio-Zabielska, A.U. The effect of high fat diet and metformin treatment on liver lipids accumulation and their impact on insulin action. Sci. Rep. 2018, 8, 7249. [Google Scholar] [CrossRef] [Green Version]
- Chocian, G.; Chabowski, A.; Żendzian-Piotrowska, M.; Harasim, E.; Łukaszuk, B.; Górski, J. High fat diet induces ceramide and sphingomyelin formation in rat’s liver nuclei. Mol. Cell. Biochem. 2010, 340, 125–131. [Google Scholar] [CrossRef] [PubMed]
- Puri, P.; Baillie, R.A.; Wiest, M.M.; Mirshahi, F.; Choudhury, J.; Cheung, O.; Sargeant, C.; Contos, M.J.; Sanyal, A.J. A lipidomic analysis of nonalcoholic fatty liver disease. Hepatology 2007, 46, 1081–1090. [Google Scholar] [CrossRef] [PubMed]
- Utzschneider, K.M.; Kahn, S.E. The Role of Insulin Resistance in Nonalcoholic Fatty Liver Disease. J. Clin. Endocrinol. Metab. 2006, 91, 4753–4761. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tardif, N.; Salles, J.; Guillet, C.; Tordjman, J.; Reggio, S.; Landrier, J.; Giraudet, C.; Patrac, V.; Bertrand-Michel, J.; Migne, C.; et al. Muscle ectopic fat deposition contributes to anabolic resistance in obese sarcopenic old rats through e IF 2α activation. Aging Cell 2014, 13, 1001–1011. [Google Scholar] [CrossRef]
- Aurich, A.-C.; Niemann, B.; Pan, R.; Gruenler, S.; Issa, H.; Silber, R.-E.; Rohrbach, S. Age-dependent effects of high fat-diet on murine left ventricles: Role of palmitate. Basic Res. Cardiol. 2013, 108, 369. [Google Scholar] [CrossRef]
- Kurek, K.; Piotrowska, D.M.; Wiesiołek-Kurek, P.; Lukaszuk, B.; Chabowski, A.; Górski, J.; Żendzian-Piotrowska, M. Inhibition of ceramide de novo synthesis reduces liver lipid accumulation in rats with nonalcoholic fatty liver disease. Liver Int. 2013, 34, 1074–1083. [Google Scholar] [CrossRef]
- Zabielski, P.; Baranowski, M.; Błachnio-Zabielska, A.; Żendzian-Piotrowska, M.; Górski, J. The effect of high-fat diet on the sphingolipid pathway of signal transduction in regenerating rat liver. Prostaglandins Other Lipid Mediat. 2010, 93, 75–83. [Google Scholar] [CrossRef] [PubMed]
- Musso, G.; Cassader, M.; Paschetta, E.; Gambino, R. Bioactive Lipid Species and Metabolic Pathways in Progression and Resolution of Nonalcoholic Steatohepatitis. Gastroenterology 2018, 155, 282–302.e8. [Google Scholar] [CrossRef]
- Kitatani, K.; Idkowiak-Baldys, J.; Hannun, Y.A. The sphingolipid salvage pathway in ceramide metabolism and signaling. Cell. Signal. 2008, 20, 1010–1018. [Google Scholar] [CrossRef] [Green Version]
- Rodriguez-Cuenca, S.; Pellegrinelli, V.; Campbell, M.; Oresic, M.; Vidal-Puig, A. Sphingolipids and glycerophospholipids—The “ying and yang” of lipotoxicity in metabolic diseases. Prog. Lipid Res. 2017, 66, 14–29. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lawton, M.; Tong, M.; Silbermann, E.; Longato, L.; Jiao, P.; Mark, P.; Wands, J.R.; Xu, H.; de la Monte, S.M. Hepatic Ceramide May Mediate Brain Insulin Resistance and Neurodegeneration in Type 2 Diabetes and Non-alcoholic Steatohepatitis. J. Alzheimer’s Dis. 2009, 16, 715–729. [Google Scholar] [CrossRef] [Green Version]
- Lee, S.; Hong, I.; Kim, B.; Shim, S.; Lee, J.S.; Lee, H.; Choi, C.S.; Park, T. Activation of sphingosine kinase 2 by endoplasmic reticulum stress ameliorates hepatic steatosis and insulin resistance in mice. Hepatology 2015, 62, 135–146. [Google Scholar] [CrossRef] [PubMed]
- Nagahashi, M.; Takabe, K.; Liu, R.; Peng, K.; Wang, X.; Wang, Y.; Hait, N.C.; Wang, X.; Allegood, J.C.; Yamada, A.; et al. Conjugated bile acid-activated S1P receptor 2 is a key regulator of sphingosine kinase 2 and hepatic gene expression. Hepatology 2015, 61, 1216–1226. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ma, M.M.; Chen, J.L.; Wang, G.G.; Wang, H.; Lu, Y.; Li, J.F.; Yi, J.; Yuan, Y.J.; Zhang, Q.W.; Mi, J.; et al. Sphingosine kinase 1 participates in insulin signalling and regulates glucose metabolism and homeostasis in KK/Ay diabetic mice. Diabetologia 2007, 50, 891–900. [Google Scholar] [CrossRef] [Green Version]
- Hsieh, C.-T.; Chuang, J.-H.; Yang, W.-C.; Yin, Y.; Lin, Y. Ceramide inhibits insulin-stimulated Akt phosphorylation through activation of Rheb/mTORC1/S6K signaling in skeletal muscle. Cell. Signal. 2014, 26, 1400–1408. [Google Scholar] [CrossRef]
- Wang, X.-X.; Ye, T.; Li, M.; Li, X.; Qiang, O.; Tang, C.-W.; Liu, R. Effects of octreotide on hepatic glycogenesis in rats with high fat diet-induced obesity. Mol. Med. Rep. 2017, 16, 109–118. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Heydemann, A.; González-Vega, M.; Berhanu, T.K.; Mull, A.J.; Sharma, R.; Holley-Cuthrell, J. Hepatic Adaptations to a High Fat Diet in the MRL Mouse Strain are Associated with an Inefficient Oxidative Phosphorylation System. Jacobs J. Diabetes Endocrinol. 2016, 2, 13. [Google Scholar]
- Ma, Z.; Chu, L.; Liu, H.; Wang, W.; Li, J.; Yao, W.; Yi, J.; Gao, Y. Beneficial effects of paeoniflorin on non-alcoholic fatty liver disease induced by high-fat diet in rats. Sci. Rep. 2017, 7, srep44819. [Google Scholar] [CrossRef] [Green Version]
- Hajduch, E.; Lachkar, F.; Ferré, P.; Foufelle, F. Roles of Ceramides in Non-Alcoholic Fatty Liver Disease. J. Clin. Med. 2021, 10, 792. [Google Scholar] [CrossRef]
- Park, J.-W.; Park, W.-J.; Kuperman, Y.; Boura-Halfon, S.; Pewzner-Jung, Y.; Futerman, A.H. Ablation of very long acyl chain sphingolipids causes hepatic insulin resistance in mice due to altered detergent-resistant membranes. Hepatology 2013, 57, 525–532. [Google Scholar] [CrossRef]
- Harasim, E.; Stepek, T.; Konstantynowicz-Nowicka, K.; Baranowski, M.; Górski, J.; Chabowski, A. Myocardial Lipid Profiling During Time Course of High Fat Diet and its Relationship to the Expression of Fatty Acid Transporters. Cell. Physiol. Biochem. 2015, 37, 1147–1158. [Google Scholar] [CrossRef]
- Chadt, A.; Al-Hasani, H. Glucose transporters in adipose tissue, liver, and skeletal muscle in metabolic health and disease. Pflügers Archiv.—Eur. J. Physiol. 2020, 472, 1273–1298. [Google Scholar] [CrossRef] [PubMed]
- Yang, Z.; Xie, C.; Xu, W.; Liu, G.; Cao, X.; Li, W.; Chen, J.; Zhu, Y.; Luo, S.; Luo, Z.; et al. Phosphorylation and inactivation of PTEN at residues Ser380/Thr382/383 induced by Helicobacter pylori promotes gastric epithelial cell survival through PI3K/Akt pathway. Oncotarget 2015, 6, 31916–31926. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Zhen, Y.-F.; Zhang, Y.-J.; Zhao, H.; Ma, H.-J.; Song, G.-Y. MicroRNA-802 regulates hepatic insulin sensitivity and glucose metabolism. Int. J. Clin. Exp. Pathol. 2018, 11, 2440–2449. [Google Scholar] [PubMed]
- Alisi, A.; Pastore, A.; Ceccarelli, S.; Panera, N.; Gnani, D.; Bruscalupi, G.; Massimi, M.; Tozzi, G.; Piemonte, F.; Nobili, V. Emodin Prevents Intrahepatic Fat Accumulation, Inflammation and Redox Status Imbalance During Diet-Induced Hepatosteatosis in Rats. Int. J. Mol. Sci. 2012, 13, 2276–2289. [Google Scholar] [CrossRef]
- Kohli, R.; Pan, X.; Malladi, P.; Wainwright, M.S.; Whitington, P.F. Mitochondrial Reactive Oxygen Species Signal Hepatocyte Steatosis by Regulating the Phosphatidylinositol 3-Kinase Cell Survival Pathway. J. Biol. Chem. 2007, 282, 21327–21336. [Google Scholar] [CrossRef] [Green Version]
- Merrill, A.; van Echten, G.; Wang, E.; Sandhoff, K. Fumonisin B1 inhibits sphingosine (sphinganine) N-acyltransferase and de novo sphingolipid biosynthesis in cultured neurons in situ. J. Biol. Chem. 1993, 268, 27299–27306. [Google Scholar] [CrossRef]
- Wang, X.; Wang, Y.; Shi, M.; Fu, H.; Xu, H.; Wei, J.; Wang, T. Mammalian target of the rapamycin pathway is involved in non-alcoholic fatty liver disease. Mol. Med. Rep. 2010, 3, 909–915. [Google Scholar] [CrossRef] [Green Version]
- Khamzina, L.; Veilleux, A.; Bergeron, S.; Marette, A. Increased Activation of the Mammalian Target of Rapamycin Pathway in Liver and Skeletal Muscle of Obese Rats: Possible Involvement in Obesity-Linked Insulin Resistance. Endocrinology 2005, 146, 1473–1481. [Google Scholar] [CrossRef] [Green Version]
- Volarević, S.; Stewart, M.J.; Ledermann, B.; Zilberman, F.; Terracciano, L.; Montini, E.; Grompe, M.; Kozma, S.C.; Thomas, G. Proliferation, But Not Growth, Blocked by Conditional Deletion of 40 S Ribosomal Protein S6. Science 2000, 288, 2045–2047. [Google Scholar] [CrossRef] [PubMed]
- Mok, K.W.; Mruk, D.D.; Cheng, C.Y. Regulation of Blood–Testis Barrier (BTB) Dynamics during Spermatogenesis via the “Yin” and “Yang” Effects of Mammalian Target of Rapamycin Complex 1 (mTORC1) and mTORC2. Int. Rev. Cell Mol. Biol. 2013, 301, 291–358. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ruvinsky, I.; Meyuhas, O. Ribosomal protein S6 phosphorylation: From protein synthesis to cell size. Trends Biochem. Sci. 2006, 31, 342–348. [Google Scholar] [CrossRef]
- Hodun, K.; Sztolsztener, K.; Chabowski, A. Antioxidants Supplementation Reduces Ceramide Synthesis Improving the Cardiac Insulin Transduction Pathway in a Rodent Model of Obesity. Nutrients 2021, 13, 3413. [Google Scholar] [CrossRef] [PubMed]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Sztolsztener, K.; Konstantynowicz-Nowicka, K.; Harasim-Symbor, E.; Chabowski, A. Time-Dependent Changes in Hepatic Sphingolipid Accumulation and PI3K/Akt/mTOR Signaling Pathway in a Rat Model of NAFLD. Int. J. Mol. Sci. 2021, 22, 12478. https://doi.org/10.3390/ijms222212478
Sztolsztener K, Konstantynowicz-Nowicka K, Harasim-Symbor E, Chabowski A. Time-Dependent Changes in Hepatic Sphingolipid Accumulation and PI3K/Akt/mTOR Signaling Pathway in a Rat Model of NAFLD. International Journal of Molecular Sciences. 2021; 22(22):12478. https://doi.org/10.3390/ijms222212478
Chicago/Turabian StyleSztolsztener, Klaudia, Karolina Konstantynowicz-Nowicka, Ewa Harasim-Symbor, and Adrian Chabowski. 2021. "Time-Dependent Changes in Hepatic Sphingolipid Accumulation and PI3K/Akt/mTOR Signaling Pathway in a Rat Model of NAFLD" International Journal of Molecular Sciences 22, no. 22: 12478. https://doi.org/10.3390/ijms222212478