Advertisement

Pharmaceutical Research

, Volume 30, Issue 9, pp 2221–2231 | Cite as

Caloric Restriction-Mediated Induction of Lipid Metabolism Gene Expression in Liver is Enhanced by Keap1-Knockdown

  • Supriya R. Kulkarni
  • Laura E. Armstrong
  • Angela L. Slitt
Research Paper

Abstract

Purpose

CR increases fatty acid oxidation to decrease tissue lipid content. The Nuclear factor E2-related factor 2 (Nrf2)-Kelch like ECH associated Protein 1 (Keap1) pathway is an antioxidant gene regulatory pathway that has been previously investigated in weight gain. However, limited interaction of Nrf2/Keap1 and CR exists. The purpose of this study was to determine how Keap1 knockdown (Keap1-KD), which is known to increase Nrf2 activity, affects the CR response, such as weight loss, hepatic lipid decrease, and induction of fatty acid oxidation gene expression.

Methods

C57BL/6 and Keap1-KD mice were maintained on 40% CR or fed ad libitum for 6 weeks. Hepatic lipid content, lipid metabolic gene, and miRNA expression was quantified.

Results

CR lowered hepatic lipid content, and induced fatty acid oxidation gene expression to a greater degree in Keap1-KD compared to C57BL/6 mice. CR differentially altered miRNA 34a, 370, let-7b* in livers of Keap1-KD compared to C57BL/6 mice.

Conclusions

CR induced induction of fatty acid oxidation gene expression was augmented with Keap1 knockdown, which was associated with differential expression of several miRNAs implicated in fatty acid oxidation and lipid accumulation.

KEY WORDS

caloric restriction gene expression liver Nfe2l2 nuclear receptor 

Abbreviations

Acc1

Acetyl-CoA carboxylase

Acot1

Acyl-CoA thioesterase 1

AL

Ad libitum

Cpt1a

Carnitine palmitoyltransferase 1A

CR

Caloric Restriction

Fabp4

Fatty acid binding protein 4

Fas

Fatty acid synthase

FXR

Farnesoid X receptor

GCLC

Glutamate cysteine ligase catalytic subunit, and

GST

Glutathione S-Transferase

InsR

insulin receptor

Keap1

Kelch like ECH-associated Protein 1

Lxr

Liver x receptor

NAFLD

Non-alcoholic fatty liver disease

NQO1

NAD(P) H:quinone oxidoreductase

Nrf2

Nuclear factor E2-Related factor 2

Pgc-1α

Peroxisome proliferator-activated receptor gamma coactivator 1-alpha

Pparα

Peroxisome Proliferator activated receptor α

RISC

RNA induced silencing complex

Scd1

Stearoyl-CoA desaturase

Sirt1

Sirtuin1

Srebp1c

Sterol regulatory element binding protein 1c

Supplementary material

11095_2013_1138_MOESM1_ESM.docx (154 kb)
ESM 1 (DOCX 153 kb)

REFERENCES

  1. 1.
    Itoh K, Igarashi K, Hayashi N, Nishizawa M, Yamamoto M. Cloning and characterization of a novel erythroid cell-derived CNC family transcription factor heterodimerizing with the small Maf family proteins. Mol Cell Biol. 1995;15:4184–93.PubMedGoogle Scholar
  2. 2.
    Moi P, Chan K, Asunis I, Cao A, Kan YW. Isolation of NF-E2-related factor 2 (Nrf2), a NF-E2-like basic leucine zipper transcriptional activator that binds to the tandem NF-E2/AP1 repeat of the beta-globin locus control region. Proc Natl Acad Sci U S A. 1994;91:9926–30.PubMedCrossRefGoogle Scholar
  3. 3.
    Maher JM, Cheng X, Slitt AL, Dieter MZ, Klaassen CD. Induction of the multidrug resistance-associated protein family of transporters by chemical activators of receptor-mediated pathways in mouse liver. Drug Metab Dispos. 2005;33:956–62.PubMedCrossRefGoogle Scholar
  4. 4.
    Maher JM, Dieter MZ, Aleksunes LM, Slitt AL, Guo G, Tanaka Y, et al. Oxidative and electrophilic stress induces multidrug resistance-associated protein transporters via the nuclear factor-E2-related factor-2 transcriptional pathway. Hepatology. 2007;46:1597–610.PubMedCrossRefGoogle Scholar
  5. 5.
    McWalter GK, Higgins LG, McLellan LI, Henderson CJ, Song L, Thornalley PJ, et al. Transcription factor Nrf2 is essential for induction of NAD(P)H:quinone oxidoreductase 1, glutathione S-transferases, and glutamate cysteine ligase by broccoli seeds and isothiocyanates. J Nutr. 2004;134:3499S–506S.PubMedGoogle Scholar
  6. 6.
    Kay HY, Kim WD, Hwang SJ, Choi HS, Gilroy RK, Wan YJ, et al. Nrf2 inhibits LXRalpha-dependent hepatic lipogenesis by competing with FXR for acetylase binding. Antioxid Redox Signal. 2011;15:2135–2146.Google Scholar
  7. 7.
    Shin S, Wakabayashi J, Yates MS, Wakabayashi N, Dolan PM, Aja S, et al. Role of Nrf2 in prevention of high-fat diet-induced obesity by synthetic triterpenoid CDDO-imidazolide. Eur J Pharmacol. 2009;620:138–44.PubMedCrossRefGoogle Scholar
  8. 8.
    Tanaka Y, Aleksunes LM, Yeager RL, Gyamfi MA, Esterly N, Guo GL, et al. NF-E2-related factor 2 inhibits lipid accumulation and oxidative stress in mice fed a high-fat diet. J Pharmacol Exp Ther. 2008;325:655–64.PubMedCrossRefGoogle Scholar
  9. 9.
    Pearson KJ, Lewis KN, Price NL, Chang JW, Perez E, Cascajo MV, et al. Nrf2 mediates cancer protection but not prolongevity induced by caloric restriction. Proc Natl Acad Sci U S A. 2008;105:2325–30.PubMedCrossRefGoogle Scholar
  10. 10.
    Kawai Y, Garduno L, Theodore M, Yang J, Arinze IJ. Acetylation-deacetylation of the transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2) regulates its transcriptional activity and nucleocytoplasmic localization. J Biol Chem. 2011;286:7629–40.PubMedCrossRefGoogle Scholar
  11. 11.
    Chalasani N, Younossi Z, Lavine JE, Diehl AM, Brunt EM, Cusi K, et al. The diagnosis and management of non-alcoholic fatty liver disease: practice guideline by the American Gastroenterological Association, American Association for the Study of Liver Diseases, and American College of Gastroenterology. Gastroenterology. 2012;142:1592–609.PubMedCrossRefGoogle Scholar
  12. 12.
    Chalasani N, Younossi Z, Lavine JE, Diehl AM, Brunt EM, Cusi K, et al. The diagnosis and management of non-alcoholic fatty liver disease: practice guideline by the American Association for the Study of Liver Diseases, American College of Gastroenterology, and the American Gastroenterological Association. Hepatology. 2012;55:2005–23.PubMedCrossRefGoogle Scholar
  13. 13.
    Cantoand C, Auwerx J. PGC-1alpha, SIRT1 and AMPK, an energy sensing network that controls energy expenditure. Curr Opin Lipidol. 2009;20:98–105.CrossRefGoogle Scholar
  14. 14.
    Jeninga EH, Schoonjans K, Auwerx J. Reversible acetylation of PGC-1: connecting energy sensors and effectors to guarantee metabolic flexibility. Oncogene. 2010;29:4617–24.PubMedCrossRefGoogle Scholar
  15. 15.
    Rodgersand JT, Puigserver P. Fasting-dependent glucose and lipid metabolic response through hepatic sirtuin 1. Proc Natl Acad Sci U S A. 2007;104:12861–6.CrossRefGoogle Scholar
  16. 16.
    Nemoto S, Fergusson MM, Finkel T. SIRT1 functionally interacts with the metabolic regulator and transcriptional coactivator PGC-1{alpha}. J Biol Chem. 2005;280:16456–60.PubMedCrossRefGoogle Scholar
  17. 17.
    Ambros V. microRNAs: tiny regulators with great potential. Cell. 2001;107:823–6.PubMedCrossRefGoogle Scholar
  18. 18.
    Pasquinelliand AE, Ruvkun G. Control of developmental timing by micrornas and their targets. Annu Rev Cell Dev Biol. 2002;18:495–513.CrossRefGoogle Scholar
  19. 19.
    Millarand AA, Waterhouse PM. Plant and animal microRNAs: similarities and differences. Funct Integr Genomics. 2005;5:129–35.CrossRefGoogle Scholar
  20. 20.
    Ebertand MS, Sharp PA. Roles for microRNAs in conferring robustness to biological processes. Cell. 2012;149:515–24.CrossRefGoogle Scholar
  21. 21.
    Ajit SK. Circulating microRNAs as biomarkers, therapeutic targets, and signaling molecules. Sensors (Basel). 2012;12:3359–69.CrossRefGoogle Scholar
  22. 22.
    Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A. 2008;105:10513–8.PubMedCrossRefGoogle Scholar
  23. 23.
    Rottiersand V, Naar AM. MicroRNAs in metabolism and metabolic disorders. Nat Rev Mol Cell Biol. 2012;13:239–50.CrossRefGoogle Scholar
  24. 24.
    Wang XW, Heegaard NH, Orum H. MicroRNAs in liver disease. Gastroenterology. 2012;142:1431–43.PubMedCrossRefGoogle Scholar
  25. 25.
    Esau C, Davis S, Murray SF, Yu XX, Pandey SK, Pear M, et al. miR-122 regulation of lipid metabolism revealed by in vivo antisense targeting. Cell Metab. 2006;3:87–98.PubMedCrossRefGoogle Scholar
  26. 26.
    Krutzfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T, Manoharan M, et al. Silencing of microRNAs in vivo with ‘antagomirs’. Nature. 2005;438:685–9.PubMedCrossRefGoogle Scholar
  27. 27.
    Cermelli S, Ruggieri A, Marrero JA, Ioannou GN, Beretta L. Circulating microRNAs in patients with chronic hepatitis C and non-alcoholic fatty liver disease. PLoS One. 2011;6:e23937.PubMedCrossRefGoogle Scholar
  28. 28.
    Yamakuchi M, Ferlito M, Lowenstein CJ. miR-34a repression of SIRT1 regulates apoptosis. Proc Natl Acad Sci U S A. 2008;105:13421–6.PubMedCrossRefGoogle Scholar
  29. 29.
    Leeand J, Kemper JK. Controlling SIRT1 expression by microRNAs in health and metabolic disease. Aging (Albany NY). 2010;2:527–34.Google Scholar
  30. 30.
    Davalos A, Goedeke L, Smibert P, Ramirez CM, Warrier NP, Andreo U, et al. miR-33a/b contribute to the regulation of fatty acid metabolism and insulin signaling. Proc Natl Acad Sci U S A. 2011;108:9232–7.PubMedCrossRefGoogle Scholar
  31. 31.
    Rayner KJ, Suarez Y, Davalos A, Parathath S, Fitzgerald ML, Tamehiro N, et al. MiR-33 contributes to the regulation of cholesterol homeostasis. Science. 2010;328:1570–3.PubMedCrossRefGoogle Scholar
  32. 32.
    Ogawa T, Enomoto M, Fujii H, Sekiya Y, Yoshizato K, Ikeda K, et al. MicroRNA-221/222 upregulation indicates the activation of stellate cells and the progression of liver fibrosis. Gut. 2012;61:1600–9.PubMedCrossRefGoogle Scholar
  33. 33.
    Sun T, Fu M, Bookout AL, Kliewer SA, Mangelsdorf DJ. MicroRNA let-7 regulates 3T3-L1 adipogenesis. Mol Endocrinol. 2009;23:925–31.PubMedCrossRefGoogle Scholar
  34. 34.
    Zhu H, Shyh-Chang N, Segre AV, Shinoda G, Shah SP, Einhorn WS, et al. The Lin28/let-7 axis regulates glucose metabolism. Cell. 2011;147:81–94.PubMedCrossRefGoogle Scholar
  35. 35.
    Wu KC, Cui JY, Klaassen CD. Beneficial role of Nrf2 in regulating NADPH generation and consumption. Toxicol Sci. 2011;123:590–600.PubMedCrossRefGoogle Scholar
  36. 36.
    Reisman SA, Yeager RL, Yamamoto M, Klaassen CD. Increased Nrf2 activation in livers from Keap1-knockdown mice increases expression of cytoprotective genes that detoxify electrophiles more than those that detoxify reactive oxygen species. Toxicol Sci. 2009;108:35–47.PubMedCrossRefGoogle Scholar
  37. 37.
    Itoh K, Chiba T, Takahashi S, Ishii T, Igarashi K, Katoh Y, et al. An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements. Biochem Biophys Res Commun. 1997;236:313–22.PubMedCrossRefGoogle Scholar
  38. 38.
    Xu J, Kulkarni SR, Donepudi AC, More VR, Slitt AL. Enhanced Nrf2 activity worsens insulin resistance, impairs lipid accumulation in adipose tissue, and increases hepatic steatosis in leptin-deficient mice. Diabetes. 2012;61:3208–18.PubMedCrossRefGoogle Scholar
  39. 39.
    Chen D, Bruno J, Easlon E, Lin SJ, Cheng HL, Alt FW, et al. Tissue-specific regulation of SIRT1 by calorie restriction. Genes Dev. 2008;22:1753–7.PubMedCrossRefGoogle Scholar
  40. 40.
    Dongol B, Shah Y, Kim I, Gonzalez FJ, Hunt MC. The acyl-CoA thioesterase I is regulated by PPARalpha and HNF4alpha via a distal response element in the promoter. J Lipid Res. 2007;48:1781–91.PubMedCrossRefGoogle Scholar
  41. 41.
    Stulnig TM, Steffensen KR, Gao H, Reimers M, Dahlman-Wright K, Schuster GU, et al. Novel roles of liver X receptors exposed by gene expression profiling in liver and adipose tissue. Mol Pharmacol. 2002;62:1299–305.PubMedCrossRefGoogle Scholar
  42. 42.
    Zhang YK, Saupe KW, Klaassen CD. Energy restriction does not compensate for the reduced expression of hepatic drug-processing genes in mice with aging. Drug Metab Dispos. 2010;38:1122–31.PubMedCrossRefGoogle Scholar
  43. 43.
    Reisman SA, Csanaky IL, Aleksunes LM, Klaassen CD. Altered disposition of acetaminophen in Nrf2-null and Keap1-knockdown mice. Toxicol Sci. 2009;109:31–40.PubMedCrossRefGoogle Scholar
  44. 44.
    Lee, AP, Sharma A, Song G, Miao J, Mo YY, Wang L, et al. A pathway involving farnesoid X receptor and small heterodimer partner positively regulates hepatic sirtuin 1 levels via microRNA-34a inhibition. The J Biol Chem. 2010;285:12604–12611Google Scholar
  45. 45.
    Castro RE, Ferreira DM, Afonso MB, Borralho PM, Machado MV, Cortez-Pinto H, et al. miR-34a/SIRT1/p53 is suppressed by ursodeoxycholic acid in the rat liver and activated by disease severity in human non-alcoholic fatty liver disease. J Hepatol. 2013;58(1):119–25Google Scholar
  46. 46.
    More VR, Xu J, Shimpi PC, Belgrave C, Luyendyk JP, Yamamoto M, et al. Keap1 knockdown increases markers of metabolic syndrome after long-term fat diet feeding. Free Radic Biol Med. 2013.Google Scholar
  47. 47.
    Zhang YK, Yeager RL, Tanaka Y, Klaassen CD. Enhanced expression of Nrf2 in mice attenuates the fatty liver produced by a methionine- and choline-deficient diet. Toxicol Appl Pharmacol. 2010;245:326–34.PubMedCrossRefGoogle Scholar
  48. 48.
    Bouchard L, Rabasa-Lhoret R, Faraj M, Lavoie ME, Mill J, Perusse L, et al. Differential epigenomic and transcriptomic responses in subcutaneous adipose tissue between low and high responders to caloric restriction. Am J Clin Nutr. 2010;91:309–20.PubMedCrossRefGoogle Scholar
  49. 49.
    Milagro FI, Campion J, Cordero P, Goyenechea E, Gomez-Uriz AM, Abete I, et al. A dual epigenomic approach for the search of obesity biomarkers: DNA methylation in relation to diet-induced weight loss. FASEB J. 2011;25:1378–89.PubMedCrossRefGoogle Scholar
  50. 50.
    Chaudharyand N, Pfluger PT. Metabolic benefits from Sirt1 and Sirt1 activators. Curr Opin Clin Nutr Metab Care. 2009;12:431–7.CrossRefGoogle Scholar
  51. 51.
    Yamakuchi M, Lowenstein CJ. MiR-34, SIRT1 and p53: the feedback loop. Cell Cycle Georgetown, Tex. 2009;8:712–5.PubMedCrossRefGoogle Scholar
  52. 52.
    Iliopoulos D, Drosatos K, Hiyama Y, Goldberg IJ, Zannis VI. MicroRNA-370 controls the expression of microRNA-122 and Cpt1alpha and affects lipid metabolism. J Lipid Res. 2010;51:1513–23.PubMedCrossRefGoogle Scholar
  53. 53.
    Jaiswal AK. Nrf2 signaling in coordinated activation of antioxidant gene expression. Free Radic Biol Med. 2004;36:1199–207.PubMedCrossRefGoogle Scholar
  54. 54.
    Frostand RJ, Olson EN. Control of glucose homeostasis and insulin sensitivity by the Let-7 family of microRNAs. Proc Natl Acad Sci USA. 2011;108:21075–21080Google Scholar
  55. 55.
    Hou W, Tian Q, Steuerwald NM, Schrum LW, Bonkovsky HL. The let-7 microRNA enhances heme oxygenase-1 by suppressing Bach1 and attenuates oxidant injury in human hepatocytes. Biochim Biophys Acta. 2012;1819:1113–22.PubMedCrossRefGoogle Scholar
  56. 56.
    Sangokoya C, Telen MJ, Chi JT. microRNA miR-144 modulates oxidative stress tolerance and associates with anemia severity in sickle cell disease. Blood. 2010;116:4338–48.PubMedCrossRefGoogle Scholar
  57. 57.
    Wu KC, Liu J, Klaassen CD. Role of Nrf2 in preventing ethanol-induced oxidative stress and lipid accumulation. Toxicol Appl Pharmacol. 2012;262:321–329.Google Scholar
  58. 58.
    Mori MA, Raghavan P, Thomou T, Boucher J, Robida-Stubbs S, Macotela Y, et al. Role of microRNA processing in adipose tissue in stress defense and longevity. Cell Metab. 2012;16:336–47.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Supriya R. Kulkarni
    • 1
  • Laura E. Armstrong
    • 1
  • Angela L. Slitt
    • 1
  1. 1.Department of Biomedical and Pharmaceutical SciencesUniversity of Rhode IslandKingstonUSA

Personalised recommendations