Resveratrol-induced brown fat-like phenotype in 3T3-L1 adipocytes partly via mTOR pathway
Background: Browning of white adipose tissues (WAT) is recognized as a novel way to combat obesity and its related comorbidities. Thus, a lot of dietary agents contributing to browning of WAT have been identified.
Objective: In this study, we try to explore the mechanism of the browning of WAT induced by resveratrol (Res) in 3T3-L1 adipocytes.
Methods: The levels of cell viability and lipid accumulation were evaluated under different concentrations of Res. Cell signaling pathway analysis was performed to investigate the possible mechanisms of the WAT browning effect of Res in 3T3-L1 cells.
Results: We found that Res induced the brown fat-like phenotype by activating protein expressions of brown adipocyte-specific markers, such as peroxisome proliferator-activated receptor gamma (PPAR-γ), peroxisome proliferator- activated receptor gamma coactivator-1 alpha (PGC-1α), and uncoupling protein 1 (UCP1). Besides, Res reduced lipid accumulation, as shown by Oil Red O staining. The increased small lipid droplets implied that Res-treated 3T3-L1 adipocytes had some features of brown adipocytes. The brown fat-like phenotype in 3T3-L1 adipocytes induced by Res was possibly mediated by activation of mammalian target of rapamycin (mTOR), as brown adipocyte-specific markers were decreased by rapamycin, an inhibitor of mTOR and the MHY1485 treatment, an activator of mTOR, showed the similar effect of Res on browning markers.
Conclusions: Res induced brown-like adipocyte phenotype in 3T3-L1 adipocytes partly via mTOR pathway, which provided new insights into the utilization of Res to prevent obesity and related comorbidities.
- Afshin A, Forouzanfar MH, Reitsma MB, Sur P, Estep K, Lee A, et al. Health effects of overweight and obesity in 195 countries over 25 years. N Engl J Med 2017; 377: 13–27. doi: 10.1056/NEJMoa1614362
- Cypess AM, Kahn CR. Brown fat as a therapy for obesity and diabetes. Curr Opin Endocrinol Diabetes Obes 2010; 17: 143–149. doi: 10.1097/MED.0b013e328337a81f
- Wu J, Bostrom P, Sparks LM, Ye L, Choi JH, Giang A.-H, et al. Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell 2012; 150: 366–376. doi: 10.1016/j.cell.2012.05.016
- Lone J, Choi JH, Kim SW, Yun JW. Curcumin induces brown fat-like phenotype in 3T3-L1 and primary white adipocytes. J Nutr Biochem 2016; 27: 193–202. doi: 10.1016/j.jnutbio.2015.09.006
- Choi JH, Yun JW. Chrysin induces brown fat-like phenotype and enhances lipid metabolism in 3T3-L1 adipocytes. Nutrition 2016; 32: 1002–1010. doi: 10.1016/j.nut.2016.02.007
- Baskaran P, Krishnan V, Ren J, Thyagarajan B. Capsaicin induces browning of white adipose tissue and counters obesity by activating TRPV1 channel-dependent mechanisms. Br J Pharmacol 2016; 173: 2369–2389. doi: 10.1111/bph.13514
- Hsu H.-T, Tseng Y.-T, Wong W.-J, Liu C.-M, Lo Y.-C. Resveratrol prevents nanoparticles-induced inflammation and oxidative stress via downregulation of PKC-alpha and NADPH oxidase in lung epithelial A549 cells. BMC Complement Altern Med 2018; 18: 211. doi: 10.1186/s12906-018-2278-6
- Chatterjee K, Mukherjee S, Vanmanen J, Banerjee P, Fata JE. Dietary polyphenols, resveratrol and pterostilbene exhibit antitumor activity on an HPV E6-positive cervical cancer model: an in vitro and in vivo analysis. Front Oncol 2019; 9: 352. doi: 10.3389/fonc.2019.00352
- Aguirre L, Fernandez-Quintela A, Arias N, Portillo MP. Resveratrol: anti-obesity mechanisms of action. Molecules (Basel, Switzerland) 2014; 19: 18632–18655. doi: 10.3390/molecules191118632
- Arias N, Pico C, Macarulla MT, Oliver P, Miranda J, Palou A, et al. A combination of resveratrol and quercetin induces browning in white adipose tissue of rats fed an obesogenic diet. Obesity 2017; 25: 111–121. doi: 10.1002/oby.21706
- Andrade JM, Frade AC, Guimaraes JB, Freitas KM, Lopes MT, Guimaraes AL, et al. Resveratrol increases brown adipose tissue thermogenesis markers by increasing SIRT1 and energy expenditure and decreasing fat accumulation in adipose tissue of mice fed a standard diet. Eur J Nutr 2014; 53: 1503–1510. doi: 10.1007/s00394-014-0655-6
- Ravaud C, Pare M, Yao X, Azoulay S, Mazure NM, Dani C, et al. Resveratrol and HIV-protease inhibitors control UCP1 expression through opposite effects on p38 MAPK phosphorylation in human adipocytes. J Cell Physiol 2019; 235: 1184–1196. doi: 10.1002/jcp.29032
- Andrade JMO, Barcala-Jorge AS, Batista-Jorge GC, Paraiso AF, Freitas KM, Lelis DF, et al. Effect of resveratrol on expression of genes involved thermogenesis in mice and humans. Biomed Pharmacother 2019; 112: 108634. doi: 10.1016/j.biopha.2019.108634
- Li Q, Wang K, Ma Y, Qin C, Dong C, Jin P, et al. Resveratrol derivative BTM-0512 mitigates obesity by promoting beige remodeling of subcutaneous preadipocytes. Acta Biochim Et Biophys Sin 2017; 49: 318–327. doi: 10.1093/abbs/gmx009
- Lee PL, Jung SM, Guertin DA. The complex roles of mechanistic target of rapamycin in adipocytes and beyond. Trends Endocrinol Metab 2017; 28: 319–339. doi: 10.1016/j.tem.2017.01.004
- Leipert J, Kaessner F, Schuster S, Haendel N, Koerner A, Kiess W, et al. Resveratrol potentiates growth inhibitory effects of rapamycin in PTEN-deficient lipoma cells by suppressing p70S6 kinase activity. Nutr Cancer 2016; 68: 342–349. doi: 10.1080/01635581.2016.1145244
- Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65: 55–63. doi: 10.1016/0022-1759(83)90303-4
- Cheng Y, Prusoff WH. Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol 1973; 22: 3099–3108. doi: 10.1016/0006-2952(73)90196-2
- Wang S, Liang X, Yang Q, Fu X, Rogers CJ, Zhu M, et al. Resveratrol induces brown-like adipocyte formation in white fat through activation of AMP-activated protein kinase (AMPK) alpha 1. Int J Obesity 2015; 39: 967–976. doi: 10.1038/ijo.2015.23
- Shabalina IG, Petrovic N, de Jong JMA, Kalinovich AV, Cannon B, Nedergaard J. UCP1 in brite/beige adipose tissue mitochondria is functionally thermogenic. Cell Rep 2013; 5: 1196–1203. doi: 10.1016/j.celrep.2013.10.044
- Wang H, Liu L, Lin JZ, Aprahamian TR, Farmer SR. Browning of white adipose tissue with roscovitine induces a distinct population of UCP1(+) adipocytes. Cell Metab 2016; 24: 835–847. doi: 10.1016/j.cmet.2016.10.005
- Lo KA, Sun L. Turning WAT into BAT: a review on regulators controlling the browning of white adipocytes. Biosci Rep 2013; 33: 711–719. doi: 10.1042/bsr20130046
- Villarroya F, Peyrou M, Giralt M. Transcriptional regulation of the uncoupling protein-1 gene. Biochimie 2017; 134: 86–92. doi: 10.1016/j.biochi.2016.09.017
- Shan T, Zhang P, Jiang Q, Xiong Y, Wang Y, Kuang S. Adipocyte-specific deletion of mTOR inhibits adipose tissue development and causes insulin resistance in mice. Diabetologia 2016; 59: 1995–2004. doi: 10.1007/s00125-016-4006-4
- Polak P, Cybulski N, Feige JN, Auwerx J, Ruegg MA, Hall MN. Adipose-specific knockout of raptor results in lean mice with enhanced mitochondrial respiration. Cell Metab 2008; 8: 399–410. doi: 10.1016/j.cmet.2008.09.003
- Liu D, Bordicchia M, Zhang C, Fang H, Wei W, Li J.-L, et al. Activation of mTORC1 is essential for beta-adrenergic stimulation of adipose browning. J Clin Invest 2016; 126: 1704–1716. doi: 10.1172/jci83532
- Tran CM, Mukherjee S, Ye L, Frederick DW, Kissig M, Davis JG, et al. Rapamycin blocks induction of the thermogenic program in white adipose tissue. Diabetes 2016; 65: 927–941. doi: 10.2337/db15-0502
- Dolinsky VW, Chan AYM, Frayne IR, Light PE, Rosiers CD, Dyck JRB. Resveratrol prevents the prohypertrophic effects of oxidative stress on LKB1. Circulation 2009; 119: 1643–1652. doi: 10.1161/circulationaha.108.787440
- Rajapakse AG, Yepuri G, Carvas JM, Stein S, Matter CM, Scerri I, et al. Hyperactive S6K1 mediates oxidative stress and endothelial dysfunction in aging: inhibition by resveratrol. PLoS One 2011; 6: e19237. doi: 10.1371/journal.pone.0019237
- Demidenko ZN, Blagosklonny MV. At concentrations that inhibit mTOR, resveratrol suppresses cellular senescence. Cell Cycle 2009; 8: 1901–1904. doi: 10.4161/cc.8.12.8810
- Li N, Du Z, Shen Q, Lei Q, Zhang Y, Zhang M, et al. Resveratrol enhances self-renewal of mouse embryonic stem cells. J Cell Biochem 2017; 118: 1928–1935. doi: 10.1002/jcb.25942
- Liu M, Liu F. Resveratrol inhibits mTOR signaling by targeting DEPTOR. Commun Integr Biol 2011; 4: 382–384. doi: 10.4161/cib.4.4.15309
- Armour SM, Baur JA, Hsieh SN, Land-Bracha A, Thomas SM, Sinclair DA. Inhibition of mammalian S6 kinase by resveratrol suppresses autophagy. Aging 2009; 1: 515–528. doi: 10.18632/aging.100056
- Huang J, Gan Q, Han L, Li J, Zhang H, Sun Y, et al. SIRT1 overexpression antagonizes cellular senescence with activated ERK/S6k1 signaling in human diploid fibroblasts. PLoS One 2008; 3: e1710. doi: 10.1371/journal.pone.0001710
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