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Activation of SIRT1 by l-serine increases fatty acid oxidation and reverses insulin resistance in C2C12 myotubes (l-serine activates SIRT1 in C2C12 myotubes)

  • Woo-Cheol Sim
  • Dong Gwang Kim
  • Wonseok Lee
  • Hyungtai Sim
  • You-Jin Choi
  • Byung-Hoon LeeEmail author
Original Article

Abstract

Silent information regulator 1 (SIRT1) is a nicotinamide adenine dinucleotide (NAD+)-dependent deacetylase, and the function is linked to cellular metabolism including mitochondrial biogenesis. Hepatic l-serine concentration is decreased significantly in fatty liver disease. We reported that the supplementation of the amino acid ameliorated the alcoholic fatty liver by enhancing l-serine-dependent homocysteine metabolism. In this study, we hypothesized that the metabolic production of NAD+ from l-serine and thus activation of SIRT1 contribute to the action of l-serine. To this end, we evaluated the effects of l-serine on SIRT1 activity and mitochondria biogenesis in C2C12 myotubes. l-Serine increased intracellular NAD+ content and led to the activation of SIRT1 as determined by p53 luciferase assay and western blot analysis of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) acetylation. l-Serine treatment increased the expression of the genes associated with mitochondrial biogenesis and enhanced mitochondrial mass and function. In addition, l-serine reversed cellular insulin resistance determined by insulin-induced phosphorylation of Akt and GLUT4 expression and membrane translocation. l-Serine-induced mitochondrial gene expression, fatty acid oxidation, and insulin sensitization were mediated by enhanced SIRT1 activity, which was verified by selective SIRT1 inhibitor (Ex-527) and siRNA directed to SIRT1. l-Serine effect on cellular NAD+ level is dependent on the l-serine metabolism to pyruvate that is subsequently converted to lactate by lactate dehydrogenase. In summary, these data suggest that l-serine increases cellular NAD+ level and thus SIRT1 activity in C2C12 myotubes.

Keywords

Insulin resistance l-Serine Mitochondrial biogenesis Nicotinamide adenine dinucleotide (NAD+Silent information regulator 1 (SIRT1) 

Abbreviations

AMPK

AMP-activated protein kinase

DMEM

Dulbecco’s modified Eagle’s medium

DPBS

Dulbecco’s phosphate buffered saline

ER

Endoplasmic reticulum

GLUT4

Glucose transporter type 4

IP

Immunoprecipitation

LDH

Lactate dehydrogenase

NAD+

Nicotinamide adenine dinucleotide

NMN

Nicotinamide mononucleotide

NAMPT

Nicotinamide phosphoribosyltransferase

OCR

Oxygen consumption rate

PGC-1α

Peroxisome proliferator-activated receptor gamma coactivator 1-alpha

PHGDH

Phosphoglycerate dehydrogenase

PKM2

Pyruvate kinase M2

qRT-PCR

Quantitative real-time polymerase chain reaction

SIRT1

Silent information regulator 1

Notes

Funding information

This research was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. 2017R1A2B4003179) and a grant (16173MFDS009) from Ministry of Food and Drug Safety in 2017.

Supplementary material

10565_2019_9463_MOESM1_ESM.docx (15 kb)
Supplemental Table 1 (DOCX 15 kb)
10565_2019_9463_MOESM2_ESM.pptx (213 kb)
Supplemental Figure 1 (PPTX 213 kb)
10565_2019_9463_MOESM3_ESM.pptx (126 kb)
Supplemental Figure 2 (PPTX 126 kb)
10565_2019_9463_MOESM4_ESM.pptx (296 kb)
Supplemental Figure 3 (PPTX 295 kb)

References

  1. Bai P, Cantó C, Oudart H, Brunyánszki A, Cen Y, Thomas C, et al. PARP-1 inhibition increases mitochondrial metabolism through SIRT1 activation. Cell Metab. 2011;13:461–8.CrossRefGoogle Scholar
  2. Beaudoin MS, Snook LA, Arkell AM, Simpson JA, Holloway GP, Wright DC. Resveratrol supplementation improves white adipose tissue function in a depot-specific manner in Zucker diabetic fatty rats. Am J Phys Regul Integr Comp Phys. 2013;305:R542–51.Google Scholar
  3. Bortolotti M, Kreis R, Debard C, Cariou B, Faeh D, Chetiveaux M, et al. High protein intake reduces intrahepatocellular lipid deposition in humans. Am J Clin Nutr. 2009;90:1002–10.CrossRefGoogle Scholar
  4. Braidy N, Guillemin GJ, Grant R. Effects of kynurenine pathway inhibition on NAD metabolism and cell viability in human primary astrocytes and neurons. Int J Tryptophan Res. 2011;4:29–37.CrossRefGoogle Scholar
  5. Bruckbauer A, Zemel MB. Synergistic effects of polyphenols and methylxanthines with leucine on AMPK/Sirtuin-mediated metabolism in muscle cells and adipocytes. PLoS One. 2014;9:e89166.CrossRefGoogle Scholar
  6. Burgner JW 2nd, Ray WJ Jr. On the origin of the lactate dehydrogenase induced rate effect. Biochemistry. 1984;23:3636–48.CrossRefGoogle Scholar
  7. Cantó C, Gerhart-Hines Z, Feige JN, Lagouge M, Noriega L, Milne JC, et al. AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity. Nature. 2009;458:1056–60.CrossRefGoogle Scholar
  8. Cantó C, Houtkooper RH, Pirinen E, Youn DY, Oosterveer MH, Cen Y, et al. The NAD(+) precursor nicotinamide riboside enhances oxidative metabolism and protects against high-fat diet-induced obesity. Cell Metab. 2012;15:838–47.CrossRefGoogle Scholar
  9. Chaneton B, Hillmann P, Zheng L, Martin ACL, Maddocks ODK, Chokkathukalam A, et al. Serine is a natural ligand and allosteric activator of pyruvate kinase M2. Nature. 2012;491:458–62.CrossRefGoogle Scholar
  10. Chang HC, Guarente L. SIRT1 mediates central circadian control in the SCN by a mechanism that decays with aging. Cell. 2013;153:1448–60.CrossRefGoogle Scholar
  11. Chen G, Ma C, Bower KA, Shi X, Ke Z, Luo J. Ethanol promotes endoplasmic reticulum stress-induced neuronal death: involvement of oxidative stress. J Neurosci Res. 2008;86:937–46.CrossRefGoogle Scholar
  12. Costford SR, Bajpeyi S, Pasarica M, Albarado DC, Thomas SC, Xie H, et al. Skeletal muscle NAMPT is induced by exercise in humans. Am J Physiol Endocrinol Metab. 2010;298:E117–26.CrossRefGoogle Scholar
  13. Fröjdö S, Durand C, Molin L, Carey AL, El-Osta A, et al. Phosphoinositide 3-kinase as a novel functional target for the regulation of the insulin signaling pathway by SIRT1. Mol Cell Endocrinol. 2011;335:166–76.CrossRefGoogle Scholar
  14. Fu WJ, Haynes TE, Kohli R, Hu J, Shi W, Spencer TE, et al. Dietary L-arginine supplementation reduces fat mass in Zucker diabetic fatty rats. J Nutr. 2005;135:714–21.CrossRefGoogle Scholar
  15. Gerhart-Hines Z, Rodgers JT, Bare O, Lerin C, Kim SH, Mostoslavsky R, et al. Metabolic control of muscle mitochondrial function and fatty acid oxidation through SIRT1/PGC-1alpha. EMBO J. 2007;26:1913–23.CrossRefGoogle Scholar
  16. Gerhart-Hines Z, Dominy JE Jr, Blättler SM, Jedrychowski MP, Banks AS, Lim JH, et al. The cAMP/PKA pathway rapidly activates SIRT1 to promote fatty acid oxidation independently of changes in NAD(+). Mol Cell. 2011;44:851–63.CrossRefGoogle Scholar
  17. Gladden LB. Lactate metabolism during exercise. In: Poortmans JR, editor. Principles of exercise biochemistry. 3rd ed. Basel: Karger Press; 2004. p. 152–96.Google Scholar
  18. Iwabu M, Yamauchi T, Okada-Iwabu M, Sato K, Nakagawa T, Funata M, et al. Adiponectin and AdipoR1 regulate PGC-1alpha and mitochondria by Ca(2+) and AMPK/SIRT1. Nature. 2010;464:1313–9.CrossRefGoogle Scholar
  19. Kandel-Kfir M, Almog T, Shaish A, Shlomai G, Anafi L, Avivi C, et al. Interleukin-1α deficiency attenuates endoplasmic reticulum stress-induced liver damage and CHOP expression in mice. J Hepatol. 2015;63:926–33.CrossRefGoogle Scholar
  20. Li H, Xu M, Lee J, He C, Xie Z. Leucine supplementation increases SIRT1 expression and prevents mitochondrial dysfunction and metabolic disorders in high-fat diet-induced obese mice. Am J Physiol Endocrinol Metab. 2012;303:E1234–44.CrossRefGoogle Scholar
  21. Li Y, Wong K, Giles A, Jiang J, Lee JW, Adams AC, et al. Hepatic SIRT1 attenuates hepatic steatosis and controls energy balance in mice by inducing fibroblast growth factor 21. Gastroenterology. 2014;146:539–49.CrossRefGoogle Scholar
  22. Li HB, Yang YRY, Mo ZJ, Ding Y, Jiang WJ. Silibinin improves palmitate-induced insulin resistance in C2C12 myotubes by attenuating IRS-1/PI3K/Akt pathway inhibition. Braz J Med Biol Res. 2015;48:440–6.CrossRefGoogle Scholar
  23. Mattaini KR, Sullivan MR, Vander Heiden MG. The importance of serine metabolism in cancer. J Cell Biol. 2016;214:249–57.CrossRefGoogle Scholar
  24. Opara EC, Petro A, Tevrizian A, Feinglosk MN, Surwit RS. L-glutamine supplementation of a high fat diet reduces body weight and attenuates hyperglycemia and hyperinsulinemia in C57BL/6J mice. J Nutr. 1996;126:273–9.CrossRefGoogle Scholar
  25. Philp A, Perez-Schindler J, Green C, Hamilton DL, Baar K. Pyruvate suppresses PGC1alpha expression and substrate utilization despite increased respiratory chain content in C2C12 myotubes. Am J Phys Cell Phys. 2010;299:C240–50.CrossRefGoogle Scholar
  26. Price NL, Gomes AP, Ling AJ, Duarte FV, Martin-Montalvo A, et al. SIRT1 is required for AMPK activation and the beneficial effects of resveratrol on mitochondrial function. Cell Metab. 2012;15:675–90.CrossRefGoogle Scholar
  27. Sim WC, Yin HQ, Choi HS, Choi YJ, Kwak HC, Kim SK, et al. L-serine supplementation attenuates alcoholic fatty liver by enhancing homocysteine metabolism in mice and rats. J Nutr. 2015;145:260–7.CrossRefGoogle Scholar
  28. Sim WC, Han I, Lee W, Choi YJ, Lee KY, Kim DG, et al. Inhibition of homocysteine-induced endoplasmic reticulum stress and endothelial cell damage by l-serine and glycine. Toxicol in Vitro. 2016;34:138–45.CrossRefGoogle Scholar
  29. Singh BK, Sinha RA, Zhou J, Xie SY, You SH, Gauthier K, et al. FoxO1 deacetylation regulates thyroid hormone-induced transcription of key hepatic gluconeogenic genes. J Biol Chem. 2013;288:30365–72.CrossRefGoogle Scholar
  30. Suchankova G, Nelson LE, Gerhart-Hines Z, Kelly M, Gauthier MS, Saha AK, et al. Concurrent regulation of AMP-activated protein kinase and SIRT1 in mammalian cells. Biochem Biophys Res Commun. 2009;378:836–41.CrossRefGoogle Scholar
  31. Sun X, Zemel MB. Leucine modulation of mitochondrial mass and oxygen consumption in skeletal muscle cells and adipocytes. Nutr Metab (Lond). 2009;6:26.  https://doi.org/10.1186/1743-7075-6-26.CrossRefGoogle Scholar
  32. Sun Q, Hu H, Wang W, Jin H, Feng G, Jia N. Taurine attenuates amyloid β 1-42-induced mitochondrial dysfunction by activating of SIRT1 in SK-N-SH cells. Biochem Biophys Res Commun. 2014;447:485–9.CrossRefGoogle Scholar
  33. Timmers S, Konings E, Bilet L, Houtkooper RH, van de Weijer T, Goossens GH, et al. Calorie restriction-like effects of 30 days of resveratrol supplementation on energy metabolism and metabolic profile in obese humans. Cell Metab. 2011;14:612–22.CrossRefGoogle Scholar
  34. Tremblay F, Lavigne C, Jacques H, Marette A. Role of dietary proteins and amino acids in the pathogenesis of insulin resistance. Annu Rev Nutr. 2007;27:293–310.CrossRefGoogle Scholar
  35. Wang RH, Kim HS, Xiao C, Xu X, Gavrilova O, et al. Hepatic Sirt1 deficiency in mice impairs mTorc2/Akt signaling and results in hyperglycemia, oxidative damage, and insulin resistance. J Clin Invest. 2011;121:4470–90.Google Scholar
  36. Wilson L, Yang Q, Szustakowski JD, Gullicksen PS, Halse R. Pyruvate induces mitochondrial biogenesis by a PGC-1 alpha-independent mechanism. Am J Phys Cell Phys. 2007;292:C1599–605.CrossRefGoogle Scholar
  37. Yi HW, Ma YX, Wang XN, Wang CF, Lu J, et al. Ethanol promotes saturated fatty acid-induced hepatoxicity through endoplasmic reticulum (ER) stress response. Chin J Nat Med. 2015;13:250–6.Google Scholar
  38. Yoshino J, Mills KF, Yoon MJ, Imai S. Nicotinamide mononucleotide, a key NAD(+) intermediate, treats the pathophysiology of diet- and age-induced diabetes in mice. Cell Metab. 2011;14:528–36.CrossRefGoogle Scholar
  39. Zhou X, He L, Zuo S, Zhang Y, Wan D, Long C, et al. Serine prevented high-fat diet-induced oxidative stress by activating AMPK and epigenetically modulating the expression of glutathione synthesis-related genes. Biochim Biophys Acta. 2018;1864:488–98.CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Woo-Cheol Sim
    • 1
  • Dong Gwang Kim
    • 1
  • Wonseok Lee
    • 1
  • Hyungtai Sim
    • 1
  • You-Jin Choi
    • 1
  • Byung-Hoon Lee
    • 1
    Email author
  1. 1.College of Pharmacy and Research Institute of Pharmaceutical SciencesSeoul National UniversitySeoulRepublic of Korea

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