, Volume 62, Issue 10, pp 1915–1927 | Cite as

Synergistic activation of thermogenic adipocytes by a combination of PPARγ activation, SMAD3 inhibition and adrenergic receptor activation ameliorates metabolic abnormalities in rodents

  • Tomohiro Matsumoto
  • Satomi Kiuchi
  • Takatoshi MuraseEmail author



To treat obesity and related diseases, considerable effort has gone into developing strategies to convert white adipocytes into thermogenic brown-like adipocytes (‘browning’). The purpose of this study was to identify the most efficient signal control for browning.


To identify the most efficient signal control for browning, we examined rat stromal vascular fraction cells. In addition, physiological changes consequent to signal control were examined in vivo using lean and diet-induced obese (DIO) C57BL/6J mice.


Combined treatment with the peroxisome proliferator-activated receptor γ (PPARγ) agonist rosiglitazone, the SMAD3 inhibitor SIS3 and the adrenergic receptor agonist noradrenaline (norepinephrine) synergistically induced Ucp1, Fgf21 and Cited1 expression, triggering brown adipogenesis. Synergistic induction of Ucp1 by the three agents was negatively regulated by forkhead box O (FOXO)3 via the inhibition of PPARγ-dependent gene transcription. Moreover, the administration of rosiglitazone, SIS3 and the selective β3 adrenergic receptor agonist CL316,243 to DIO mice reduced the amount of body-fat deposits (body weight from day 0 to 14, 12.3% reduction), concomitant with morphological changes in white adipose tissue, an increase in mitochondrial biosynthesis and a marked induction of uncoupling protein 1 (UCP1). Furthermore, administration of the three agents significantly increased serum adiponectin levels (mean 65.56 μg/ml with the three agents vs 20.79 μg/ml in control mice, p < 0.05) and improved glucose and lipid tolerance.


These results suggest that the combined regulation of PPARγ, SMAD and the adrenergic receptor signalling pathway synergistically induces brown adipogenesis and may serve as an effective strategy to treat obesity and related diseases, including type 2 diabetes.


Adipocytes Browning FOXO3 Obesity PPARγ agonist 



Brown adipose tissue


C-terminal-binding protein


Diet-induced obese


Forkhead box O


Inguinal white adipose tissue


Myogenic factor 5


Non-thiazolidinedione partial agonist of peroxisome proliferator-activated receptor γ


Peroxisome proliferator-activated receptor γ coactivator 1α


Peroxisome proliferator-activated receptor γ


PRD1-BF1-RIZ1 homologous domain containing 16


Small interfering RNA


Specific inhibitor of SMAD3


Stromal vascular fraction




Uncoupling protein 1



We thank our colleagues in the Biological Science Laboratories, Kao Corporation, for the helpful discussions.

Author contributions

TMu conceived and designed the research. TMa and SK performed experiments. TMu, SK and TMa wrote the manuscript and had final approval of the version to be published. TMu is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.


This work was supported financially by Kao Corporation. The study sponsor was not involved in the design of the study; the collection, analysis or interpretation of data; writing the report; or the decision to submit the report for publication.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2019_4938_MOESM1_ESM.pdf (359 kb)
ESM 1 (PDF 359 kb)


  1. 1.
    Bray GA, Bellanger T (2006) Epidemiology, trends, and morbidities of obesity and the metabolic syndrome. Endocrine 29(1):109–117. CrossRefGoogle Scholar
  2. 2.
    Cedikova M, Kripnerová M, Dvorakova J, et al (2016) Mitochondria in white, brown, and beige adipocytes. Stem Cells Int 6067349.
  3. 3.
    Klingenberg M (1999) Uncoupling protein—a useful energy dissipator. J Bioenerg Biomembr 31(5):419–430. CrossRefGoogle Scholar
  4. 4.
    Cannon B, Nedergaard J (2004) Brown adipose tissue: function and physiological significance. Physiol Rev 84(1):277–359. CrossRefGoogle Scholar
  5. 5.
    Feldmann HM, Golozoubova V, Cannon B, Nedergaard J (2009) UCP1 ablation induces obesity and abolishes diet-induced thermogenesis in mice exempt from thermal stress by living at thermoneutrality. Cell Metab 9(2):203–209. CrossRefGoogle Scholar
  6. 6.
    Stanford KI, Middelbeek RJ, Townsend KL et al (2013) Brown adipose tissue regulates glucose homeostasis and insulin sensitivity. J Clin Invest 123(1):215–223. CrossRefGoogle Scholar
  7. 7.
    Heaton JM (1972) The distribution of brown adipose tissue in the human. J Anat 112(pt 1):35–39Google Scholar
  8. 8.
    Saito M, Okamatsu-Ogura Y, Matsushita M et al (2009) High incidence of metabolically active brown adipose tissue in healthy adult humans: effects of cold exposure and adiposity. Diabetes 58(7):1526–1531. CrossRefGoogle Scholar
  9. 9.
    Virtanen KA, Lidell ME, Orava J et al (2009) Functional brown adipose tissue in healthy adults. N Engl J Med 360(15):1518–1525. CrossRefGoogle Scholar
  10. 10.
    Yoneshiro T, Aita S, Matsushita M et al (2011) Age-related decrease in cold-activated brown adipose tissue and accumulation of body fat in healthy humans. Obesity (Silver Spring) 19(9):1755–1760. CrossRefGoogle Scholar
  11. 11.
    Yoneshiro T, Aita S, Matsushita M et al (2011) Brown adipose tissue, whole-body energy expenditure, and thermogenesis in healthy adult men. Obesity (Silver Spring) 19(1):13–16. CrossRefGoogle Scholar
  12. 12.
    van Marken Lichtenbelt WD, Vanhommerig JW, Smulders NM et al (2009) Cold-activated brown adipose tissue in healthy men. N Engl J Med 360(15):1500–1508. CrossRefGoogle Scholar
  13. 13.
    Wu J, Boström P, Sparks LM et al (2012) Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell 150(2):366–376. CrossRefGoogle Scholar
  14. 14.
    Harms M, Seale P (2013) Brown and beige fat: development, function and therapeutic potential. Nat Med 19(10):1252–1263. CrossRefGoogle Scholar
  15. 15.
    Sharp LZ, Shinoda K, Ohno H et al (2012) Human BAT possesses molecular signatures that resemble beige/brite cells. PLoS One 7(11):e49452. CrossRefGoogle Scholar
  16. 16.
    Petrovic N, Walden TB, Shabalina IG, Timmons JA, Cannon B, Nedergaard J (2010) Chronic peroxisome proliferator-activated receptor γ (PPARγ) activation of epididymally derived white adipocyte cultures reveals a population of thermogenically competent, UCP1-containing adipocytes molecularly distinct from classic brown adipocytes. J Biol Chem 285(10):7153–7164. CrossRefGoogle Scholar
  17. 17.
    Rong JX, Qiu Y, Hansen MK et al (2007) Adipose mitochondrial biogenesis is suppressed in db/db and high-fat diet-fed mice and improved by rosiglitazone. Diabetes 56(7):1751–1760. CrossRefGoogle Scholar
  18. 18.
    Kajimura S, Seale P, Tomaru T et al (2008) Regulation of the brown and white fat gene programs through a PRDM16/CtBP transcriptional complex. Genes Dev 22(10):1397–1409. CrossRefGoogle Scholar
  19. 19.
    Yoshida H, Kanamori Y, Asano H et al (2013) Regulation of brown adipogenesis by the Tgf-β family: involvement of Srebp1c in Tgf-β- and activin-induced inhibition of adipogenesis. Biochim Biophys Acta 1830(11):5027–5035. CrossRefGoogle Scholar
  20. 20.
    Tseng YH, Kokkotou E, Schulz TJ et al (2008) New role of bone morphogenetic protein 7 in brown adipogenesis and energy expenditure. Nature 454(7207):1000–1004. CrossRefGoogle Scholar
  21. 21.
    Yadav H, Quijano C, Kamaraju AK et al (2011) Protection from obesity and diabetes by blockade of TGF-β/Smad3 signaling. Cell Metab 14(1):67–79. CrossRefGoogle Scholar
  22. 22.
    Park JH, Hur W, Lee SB (2015) Intricate transcriptional networks of classical brown and beige fat cells. Front Endocrinol (Lausanne) 6:124. CrossRefGoogle Scholar
  23. 23.
    Song NJ, Chang SH, Li DY, Villanueva CJ, Park KW (2017) Induction of thermogenic adipocytes: molecular targets and thermogenic small molecules. Exp Mol Med 49(7):e353. CrossRefGoogle Scholar
  24. 24.
    Wilding J (2006) Thiazolidinediones, insulin resistance and obesity: finding a balance. Int J Clin Pract 60(10):1272–1280. CrossRefGoogle Scholar
  25. 25.
    Péronnet F, Massicotte D (1991) Table of nonprotein respiratory quotient: an update. Can J Sport Sci 16(1):23–29Google Scholar
  26. 26.
    Shimotoyodome A, Fukuoka D, Suzuki J et al (2009) Coingestion of acylglycerols differentially affects glucose-induced insulin secretion via glucose-dependent insulinotropic polypeptide in C57BL/6J mice. Endocrinology 150(5):2118–2126. CrossRefGoogle Scholar
  27. 27.
    Murase T, Yokoi Y, Misawa K et al (2012) Coffee polyphenols modulate whole-body substrate oxidation and suppress postprandial hyperglycaemia, hyperinsulinaemia and hyperlipidaemia. Br J Nutr 107(12):1757–1765. CrossRefGoogle Scholar
  28. 28.
    Onuma H, Vender Kooi BT, Boustead JN, Oeser JK, O’Brien RM (2006) Correlation between FOXO1 (FKHR) and FOXO3a (FKHRL1) binding and the inhibition of basal glucose-6-phosphatase catalytic subunit gene transcription by insulin. Mol Endocrinol 20(11):2831–2847. CrossRefGoogle Scholar
  29. 29.
    Munekata K, Sakamoto K (2009) Forkhead transcription factor Foxo1 is essential for adipocyte differentiation. In Vitro Cell Dev Biol Anim 45(10):642–651. CrossRefGoogle Scholar
  30. 30.
    MacPherson RE, Castellani L, Beaudoin MS, Wright DC (2014) Evidence for fatty acids mediating CL 316,243-induced reductions in blood glucose in mice. Am J Physiol Endocrinol Metab 307(7):E563–E570. CrossRefGoogle Scholar
  31. 31.
    Ohno H, Shinoda K, Spiegelman BM, Kajimura S (2012) PPARγ agonists induce a white-to-brown fat conversion through stabilization of PRDM16 protein. Cell Metab 15(3):395–404. CrossRefGoogle Scholar
  32. 32.
    Feng XH, Derynck R (2005) Specificity and versatility in TGF-β signaling through SMADs. Annu Rev Cell Dev Biol 21(1):659–693. CrossRefGoogle Scholar
  33. 33.
    Fan W, Imamura T, Sonoda N et al (2009) FOXO1 transrepresses peroxisome proliferator-activated receptor gamma transactivation, coordinating an insulin-induced feed-forward response in adipocytes. J Biol Chem 284(18):12188–12197. CrossRefGoogle Scholar
  34. 34.
    Tzivion G, Dobson M, Ramakrishnan G (2011) FoxO transcription factors; regulation by AKT and 14-3-3 proteins. Biochim Biophys Acta 1813(11):1938–1945. CrossRefGoogle Scholar
  35. 35.
    Nakae J, Oki M, Cao Y (2008) The FoxO transcription factors and metabolic regulation. FEBS Lett 582(1):54–67. CrossRefGoogle Scholar
  36. 36.
    Ventura-Clapier R, Garnier A, Veksler V (2008) Transcriptional control of mitochondrial biogenesis: the central role of PGC-1α. Cardiovasc Res 79(2):208–217. CrossRefGoogle Scholar
  37. 37.
    Altshuler-Keylin S, Shinoda K, Hasegawa Y et al (2016) Beige adipocyte maintenance is regulated by autophagy-induced mitochondrial clearance. Cell Metab 24(3):402–419. CrossRefGoogle Scholar
  38. 38.
    Webb AE, Brunet A (2014) FOXO transcription factors: key regulators of cellular quality control. Trends Biochem Sci 39(4):159–169. CrossRefGoogle Scholar
  39. 39.
    Furuyama T, Nakazawa T, Nakano I, Mori N (2000) Identification of the differential distribution patterns of mRNAs and consensus binding sequences for mouse DAF-16 homologues. Biochem J 349(pt 2):629–634. CrossRefGoogle Scholar
  40. 40.
    Bartelt A, Bruns OT, Reimer R et al (2011) Brown adipose tissue activity controls triglyceride clearance. Nat Med 17(2):200–205. CrossRefGoogle Scholar
  41. 41.
    Bartelt A, Widenmaier SB, Schlein C et al (2018) Brown adipose tissue thermogenic adaptation requires Nrf1-mediated proteasomal activity. Nat Med 24(3):292–303. CrossRefGoogle Scholar
  42. 42.
    Ruan H, Dong LQ (2016) Adiponectin signaling and function in insulin target tissues. J Mol Cell Biol 8(2):101–109. CrossRefGoogle Scholar
  43. 43.
    Zhu Y, Richardson JA, Parada LF, Graff JM (1998) Smad3 mutant mice develop metastatic colorectal cancer. Cell 94(6):703–714. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Biological Science LaboratoriesKao CorporationTochigiJapan

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