Journal of Food and Nutrition Research
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Journal of Food and Nutrition Research. 2021, 9(3), 163-169
DOI: 10.12691/jfnr-9-3-9
Open AccessArticle

Mechanism of Formononetin-induced Stimulation of Adipocyte Fatty Acid Oxidation and Preadipocyte Differentiation

Seok-Yeong Yu1, Youngmin Choi1, 2, Young-In Kwon3, Ok-Hwan Lee4 and Young-Cheul Kim1,

1Department of Nutrition, University of Massachusetts, Amherst, MA 01003

2National Academy of Agricultural Science, Rural Development Administration, Jeonju, Korea

3Department of Food and Nutrition, Hannam University, Daejeon, 34054 Korea

4Department of Food Science and Biotechnology, Kangwon National University, Chuncheon, Korea

Pub. Date: March 25, 2021

Cite this paper:
Seok-Yeong Yu, Youngmin Choi, Young-In Kwon, Ok-Hwan Lee and Young-Cheul Kim. Mechanism of Formononetin-induced Stimulation of Adipocyte Fatty Acid Oxidation and Preadipocyte Differentiation. Journal of Food and Nutrition Research. 2021; 9(3):163-169. doi: 10.12691/jfnr-9-3-9


Decreased adipocyte fatty acid oxidation (FAO) and impaired preadipocyte differentiation characterize hypertrophic expansion of adipose tissue (AT) from obese and insulin resistant humans and are recognized as potential mechanisms for obesity-mediated dyslipidemia. Supplementation of formononetin (FMN), one of the principal isoflavones extracted from red clover or Huangqi (Astragalus roots), has been shown to have beneficial effects on obesity-related hyperlipidemia, a well-established cardiovascular risk factor. However, a target tissue and underlying mechanism(s) through which FMN acts have been under-investigated. Thus, we investigated whether FMN promotes adipocyte FAO and preadipocyte differentiation using 3T3-L1 preadipocytes to provide potential mechanisms of FMN action. We further extended this to the culture of 10T1/2 mesenchymal stem cells (MSCs) as well as mouse AT explants to reflect in vivo effects of FMN. In fully differentiated 3T3-L1 adipocytes, FMN-treatment significantly increased the expression levels of FAO-related proteins such as pAMPK, pACC, and CPT1, all of which were consistently upregulated in AT explant cultures treated with 10 μM FMN. In addition, FMN significantly enhanced the degree of differentiation of both 3T3-L1 preadipocytes and 10T1/2 MSCs into adipocytes as evidenced by Oil Red O staining of cellular lipids. This observation correlated with increased expression levels of key adipogenic transcription factors (PPARγ and C/EBPα) and their down-stream target proteins (FABP4, Glut4 and adiponectin). Moreover, FMN failed to exert its stimulatory effects on preadipocyte differentiation in both cell types in the presence of a PPARγ antagonist, suggesting a PPARγ-dependent effect of FMN. Collectively, these data provide possible mechanisms of action of FMN on lipid metabolism and further support the favorable in vivo effects of FMN in diet and obesity-induced dyslipidemia.

3T3-L1 preadipocyte 10T1/2 mesenchymal stem cell formononetin fatty acid oxidation adipocyte differentiation

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[1]  Cignarelli, A., Genchi, V.A., Perrini, S., Natalicchio, A., Laviola, L., Giorgino, F. “Insulin and insulin receptors in adipose tissue development”. Int J Mol Sci. 20 (3). 1-20. Feb.2019.
[2]  Bays, H.E., Toth, P.P., Kris-Etherton, P.M., Abate, N., Aronne, L.J., Brown, W.V., et al. “Obesity, adiposity, and dyslipidemia: A consensus statement from the National Lipid Association.” J Clin Lipidol. 7 (3). 304-383. Jul-Aug.2013.
[3]  Marini, M.A., Succurro, E., Frontoni, S., Hribal, M.L., Andreozzi, F., Lauro, R., et al. “Metabolically healthy but obese women have an intermediate cardiovascular risk profile between healthy nonobese women and obese insulin-resistant women.” Diabetes Care. 30 (8). 2145-7 Aug.2007.
[4]  Stefan, N., Kantartzis, K., Machann, J., Schick, F., Thamer, C., Rittig, K., et al. “Identification and characterization of metabolically benign obesity in humans.” Arch Intern Med. 168 (15). 1609-16 Aug.2008.
[5]  Isakson, P., Hammarstedt, A., Gustafson, B., Smith, U. “Impaired preadipocyte differentiation in human abdominal obesity: Role of Wnt, tumor necrosis factor-α, and inflammation.” Diabetes. 58 (7). 1550-7. Jul.2009.
[6]  Martínez-Jiménez, V., Cortez-Espinosa, N., Rodríguez-Varela, E., Vega-Cárdenas, M., Briones-Espinoza, M., Ruíz-Rodríguez, V.M., et al. “Altered levels of sirtuin genes (SIRT1, SIRT2, SIRT3 and SIRT6) and their target genes in adipose tissue from individual with obesity.” Diabetes Metab Syndr. 13 (1). 582-589. Jan-Feb.2019.
[7]  Rydén, M., Andersson, D.P., Bernard, S., Spalding, K., Arner, P. “Adipocyte triglyceride turnover and lipolysis in lean and overweight subjects.” J Lipid Res. 54 (10). 2909–2913 Oct.2013.
[8]  Frayn, K., Bernard, S., Spalding, K., Arner, P. “Adipocyte triglyceride turnover is independently associated with atherogenic dyslipidemia.” J Am Heart Assoc. 1 (6). e003467. Dec.2012.
[9]  Ntambi, J.M., Kim, Y.C. “Adipocyte Differentiation and Gene Expression.” J Nutr. 130 (12). 3122S-3126S. Dec.2000.
[10]  Haberbosch W. “Effects of thiazolidinediones on dyslipidemia in patients with type 2 diabetes. Are all equally vasoprotective?.” Herz. 32 (1). 51-57. Nov.2007.
[11]  Goossens, G.H., Blaak, E.E. “Adipose Tissue Dysfunction and Impaired Metabolic Health in Human Obesity: A Matter of Oxygen?.” Front Endocrinol (Lausanne). 6. 55. Apr.2015.
[12]  Kim, J.I., Huh, J.Y., Sohn, J.H., Choe, S.S., Lee, Y.S., Lim, C.Y., et al. “Lipid-Overloaded Enlarged Adipocytes Provoke Insulin Resistance Independent of Inflammation.” Mol Cell Biol. 35 (10). 1686–1699. May.2015.
[13]  Schöttl, T., Kappler, L., Fromme, T., Klingenspor, M. “Limited OXPHOS capacity in white adipocytes is a hallmark of obesity in laboratory mice irrespective of the glucose tolerance status.” Mol Metab. 4 (9). 631-42. Jul.2015.
[14]  Choo, H.J., Kim, J.H., Kwon, O.B., Lee, C.S., Mun, J.Y., Han, S.S., et al. “Mitochondria are impaired in the adipocytes of type 2 diabetic mice.” Diabetologia. 49 (4). 784-91. Apr.2006.
[15]  Mottillo, E.P., Desjardins, E.M., Crane, J.D., Smith, B.K., Green, A.E., Ducommun, S., et al. “Lack of Adipocyte AMPK Exacerbates Insulin Resistance and Hepatic Steatosis through Brown and Beige Adipose Tissue Function.” Cell Metab. 24 (1). 118-29. Jul.2016.
[16]  Lihn, A.S., Jessen, N., Pedersen, S.B., Lund, S., Richelsen, B. “AICAR stimulates adiponectin and inhibits cytokines in adipose tissue.” Biochem Biophys Res Commun. 316 (3). 853-8. Apr.2004.
[17]  Arner, P., Bernard, S., Salehpour, M., Possnert, G., Liebl, J., Steier, P., et al. “Dynamics of human adipose lipid turnover in health and metabolic disease.” Nature. 478 (7367). 110-3. Sep.2011.
[18]  Clifton-Bligh, P.B., Nery, M.L., Clifton-Bligh, R.J., Visvalingam, S., Fulcher, G.R., Byth, K., et al. “Red clover isoflavones enriched with formononetin lower serum LDL cholesterol - A randomized, double-blind, placebo-controlled study.” Eur J Clin Nutr. 69 (1). 134-42. Jan.2015.
[19]  Gautam, J., Khedgikar, V., Kushwaha, P., Choudhary, D., Nagar, G.K., Dev, K., et al. “Formononetin, an isoflavone, activates AMP-activated protein kinase β-catenin signalling to inhibit adipogenesis and rescues C57BL/6 mice from high-fat diet-induced obesity and bone loss.” Br J Nutr. 117 (5). 645-661. Mar.2017.
[20]  Andersen, C., Kotowska, D., Tortzen, C.G., Kristiansen, K., Nielsen, J., Petersen, R.K. “2-(2-Bromophenyl)-formononetin and 2-heptyl-formononetin are PPARγ partial agonists and reduce lipid accumulation in 3T3-L1 adipocytes.” Bioorganic Med Chem. 22 (21). 6105-11. Nov.2014.
[21]  Luo, L.Y., Fan, M.X., Zhao, H.Y., Li, M.X., Wu, X., Gao, W.Y. “Pharmacokinetics and Bioavailability of the Isoflavones Formononetin and Ononin and Their in Vitro Absorption in Ussing Chamber and Caco-2 Cell Models”. J Agric Food Chem. 66 (11). 2917-2924. Mar.2018.
[22]  Osler, M.E., Zierath, J.R. Minireview: Adenosine 5’-monophosphate-activated protein kinase regulation of fatty acid oxidation in skeletal muscle.” Endocrinology. 149 (3). 935-941. Mar.2008.
[23]  Galic, S., Loh, K., Murray-Segal, L., Steinberg, G.R., Andrews, Z.B., Kemp, B.E. “AMPK signaling to acetyl-CoA carboxylase is required for fasting-and cold-induced appetite but not thermogenesis.” Elife. 13 (7). e32656 Feb.2018.
[24]  Tang, Q.Q., Otto, T.C., Lane, M.D. “Commitment of C3H10T1/2 pluripotent stem cells to the adipocyte lineage.” Proc Natl Acad Sci U S A. 101 (26). 9607-11. Jul.2004.
[25]  Spalding, K.L., Arner, E., Westermark, P.O., Bernard, S., Buchholz, B.A., Bergmann, O., et al. “Dynamics of fat cell turnover in humans.” Nature. 453 (7196). 783-7. Jun.2008.
[26]  Siersbæk, R., Nielsen, R., Mandrup, S. “PPARγ in adipocyte differentiation and metabolism - Novel insights from genome-wide studies.” FEBS Lett. 584 (15). 3242-9. Aug.2010.
[27]  Lessard, J., Laforest, S., Pelletier, M., Leboeuf, M., Blackburn, L., Tchernof, A. “Low abdominal subcutaneous preadipocyte adipogenesis is associated with visceral obesity, visceral adipocyte hypertrophy, and a dysmetabolic state.” Adipocyte. 3 (3). 197-205. Jul.2014.
[28]  Ranganathan, G., Unal, R., Pokrovskaya, I., Yao-Borengasser, A., Phanavanh, B., Lecka-Czernik, B., et al. “The lipogenic enzymes DGAT1, FAS, and LPL in adipose tissue: effects of obesity, insulin resistance, and TZD treatment.” J Lipid Res. 47 (11). 2444-50. Nov.2006.
[29]  Shen, P., Liu, M.H., Ng, T.Y., Chan, Y.H. Yong, E.L. Differential Effects of Isoflavones, from Astragalus Membranaceus and Pueraria Thomsonii, on the Activation of PPARα, PPARγ, and Adipocyte Differentiation In Vitro. J Nutr. 136 (4). 899-905. Apr.2006.
[30]  Nie, T., Zhao, S., Mao, L., Yang, Y., Sun, W., Lin, X., et al. “The natural compound, formononetin, extracted from Astragalus membranaceus increases adipocyte thermogenesis by modulating PPARγ activity.” Br J Pharmacol. 175 (9). 1439-1450. May.2018.
[31]  Flachs, P., Rossmeisl, M., Kuda, O., Kopecky, J. “Stimulation of mitochondrial oxidative capacity in white fat independent of UCP1: A key to lean phenotype.” Biochim Biophys Acta. 1831 (5). 986-1003. May.2013.