Increase of Adipogenesis by Ginsenoside (Rh2) in 3T3-L1 Cell via an Activation of Glucocorticoid Receptor

Abstract

Adipocyte plays an important role in lipid regulation in mammals. Understanding of adipocyte differentiation becomes a key issue for the development of anti-obesity agent. Glucocorticoids (GCs) regulate lipid metabolism through promoting lipogenesis in adipose tissue. Ginsenoside Rh2, with a similar chemical structure as GCs, shows antidiabetic, anti-inflammatory, and anticancer actions both in vivo and in vitro. However, effect of Rh2 on glucocorticoid receptor (GR) for an increase of adipogenesis like GCs remains unclear. In the present study, we employed ginsenoside Rh2 to investigate the changes in adipogenetic process of 3T3-L1, one of the widely used preadipocytes, through activating GR or not. In leuciferase assay, we found that ginsenoside Rh2 induced GRs transitivity in a way as dexamethasone, which was deleted by RU486 at concentrations sufficient to block GR. Moreover, 3T3-L1 preadipocytes were differentiated into adipocytes by adipogenic induction medium containing 0.01 to 1 μM of ginsenoside Rh2. Also, RU486 blocked this adipogenesis induced by ginsenoside Rh2 or dexamethasone. The obtained results suggest that ginsenoside Rh2 can promote preadipocytes differentiation through activating GR. This finding seems helpful for the understanding of ginsenosides in the regulation of lipid metabolism.

ginsenoside Rh2 - glucocorticoid receptor - adipogenesis - cell line

References

• 1 Fruhbeck G, Gomez-Ambrosi J, Muruzabal FJ, Burrell MA. The adipocyte: a model for integration of endocrine and metabolic signaling in energy metabolism regulation.  Am J Physiol Endocrinol Metab. 2001;  280 E827-E847
• 2 Yamauchi T KJ, Waki H, Terauchi Y, Kubota N, Hara K, Mori Y, Ide, T MK, Tsuboyama-Kasaoka N, Ezaki O, Akanuma Y, Gavrilova O, Vinson C RM, Kagechika H, Shudo K, Yoda M, Nakano Y, Tobe, K NR, Kimura S, Tomita M, Froguel P, Kadowaki T. The fat-derived hormone adiponectin reverses insulin resistance associated with both lipoatrophy and obesity.  Nat Med. 2001;  7 941-946
• 3 Gregoire FM, Smas CM, Sul HS. Understanding adipocyte differentiation.  Physiol Rev. 1998;  78 783-809
• 4 Morrison RF, Farmer SR. Role of PPAR-γ in regulating a cascade expression of cyclin-dependent kinase inhibitors, p18(INK4c) and p21(Waf1/Cip1), during adipogenesis.  J Biol Chem. 1999;  274 17088-17097
• 5 MacDougald OA, Lane MD. Transcriptional regulation of gene expression during adipocyte differentiation.  Annu Rev Biochem. 1995;  64 345-373
• 6 Dauncey MJ, White P, Burton KA, Katsumata M. Nutrition-hormone receptor-gene interactions: implications for development and disease.  Proc Nutr Soc. 2001;  60 63-72
• 7 Berthiaume M, Laplante M, Tchernof A, Deshaies Y. Metabolic action of peroxisome proliferator-activated receptor gamma agonism in rats with exogenous hypercorticosteronemia.  Int J Obes (Lond). 2007;  31 1660-1670
• 8 Yue PY, Mak NK, Cheng YK, Leung KW, Ng TB, Fan DT, Yeung, HW, Wong RN. Pharmacogenomics and the Yin/Yang actions of ginseng: anti-tumor, angiomodulating and steroid-like activities of ginsenosides.  Chinese Medicine. 2007;  2 6-26
• 9 Lee WK, Kao ST, Liu IM, Cheng JT. Ginsenoside Rh2 is one of the active principles of Panax ginseng root to improve insulin sensitivity in fructose-rich chow-fed rats.  Horm Metab Res. 2007;  39 347-354
• 10 Yin J, Zhang H, Ye J. Traditional chinese medicine in treatment of metabolic syndrome.  Endocr Metab Immune Disord Drug Targets. 2008;  8 99-111
• 11 Xie JT, Wang CZ, Wang AB, Wu J, Basila D, Yuan CS. Antihyperglycemic effects of total ginsenosides from leaves and stem of Panax ginseng.  Acta Pharmacol Sin. 2005;  26 1104-1110
• 12 Lee YN, Lee HY, Lee YM, Chung HY, Kim SI, Lee SK, Park BC, Ki KW. Involvement of glucocorticoid receptor in the induction of differentiation by ginsenosides in F9 teratocarcinoma cells.  J Steroid Biochem Mol Biol. 1998;  67 105-111
• 13 Yang JY, Della-Fera MA, Hartzell DL, Nelson-Dooley C, Hausman DB, Baile CA. Esculetin induces apoptosis and inhibits adipogenesis in 3T3-L1 cells.  Obesity (Silver Spring). 2006;  14 1691-1699
• 14 Singh R, Artaza JN, Taylor WE, Braga M, Yuan X, Gonzalez-Cadavid NF, Bhasin S. Testosterone inhibits adipogenic differentiation in 3T3-L1 cells: nuclear translocation of androgen receptor complex with beta-catenin and T-cell factor 4 may bypass canonical Wnt signaling to down-regulate adipogenic transcription factors.  Endocrinology. 2006;  147 141-154
• 15 Shieh PC, Tsao CW, Li JS, Wu HT, Wen Y J, Kou DH, Cheng JT. Role of pituitary adenylate cyclase-activating polypeptide (PACAP) in the action of ginsenoside Rh2 against beta-amyloid-induced inhibition of rat brain astrocytes.  Neurosci Lett. 2008;  434 1-5
• 16 Rosen ED. The transcriptional basis of adipocyte development.  Prostaglandins Leukot Essent Fatty Acids. 2005;  73 31-34
• 17 Cernkovich ER, Deng J, Hua K, Harp JB. Midkine is an autocrine activator of signal transducer and activator of transcription 3 in 3T3-L1 cells.  Endocrinology. 2007;  148 1598-1604
• 18 Rubin CS, Hirsch A, Fung C, Rosen OM. Development of hormone receptors and hormonal responsiveness in vitro. Insulin receptors and insulin sensitivity in the preadipocyte and adipocyte forms of 3T3-L1 cells.  J Biol Chem. 1978;  253 7570-7578
• 19 Chapman AB, Knight DM, Dieckmann BS, Ringold GM. Analysis of gene expression during differentiation of adipogenic cells in culture and hormonal control of the developmental program.  J Biol Chem. 1984;  259 15548-15555
• 20 Gaillard D, Wabitsch M, Pipy B, Negrel R. Control of terminal differentiation of adipose precursor cells by glucocorticoids.  J Lipid Res. 1991;  32 569-579
• 21 Grigoriadis AE, Heersche JN, Aubin JE. Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone-derived clonal cell population: effect of dexamethasone.  J Cell Biol. 1988;  106 2139-2151
• 22 Gregoire F, Genart C, Hauser N, Remacle C. Glucocorticoids induce a drastic inhibition of proliferation and stimulate differentiation of adult rat fat cell precursors.  Exp Cell Res. 1991;  196 270-278
• 23 Tontonoz P, Hu E, Spiegelman BM. Regulation of adipocyte gene expression and differentiation by peroxisome proliferator activated receptor gamma.  Curr Opin Genet Dev. 1995;  5 571-576
• 24 Shang W, Yang Y, Jiang B, Jin H, Zhou L, Liu S, Chen M. Ginsenoside Rb1 promotes adipogenesis in 3T3-L1 cells by enhancing PPARgamma2 and C/EBPalpha gene expression.  Life Sci. 2007;  80 618-625
• 25 Attele AS, Wu JA, Yuan CS. Ginseng pharmacology: multiple constituents and multiple actions.  Biochem Pharmacol. 1999;  58 1685-1693
• 26 Mao J, Regelson W, Kalimi M. Molecular mechanism of RU 486 action: a review.  Mol Cell Biochem. 1992;  109 1-8
• 27 Gillis CN. Panax ginseng pharmacology: a nitric oxide link?.  Biochem Pharmacol. 1997;  54 1-8
• 28 Chung SH, Choi CG, Park SH. Comparisons between white ginseng radix and rootlet for antidiabetic activity and mechanism in KKAy mice.  Arch Pharm Res. 2001;  24 214-218
• 29 Park MY, Lee KS, Sung MK. Effects of dietary mulberry, Korean red ginseng, and banaba on glucose homeostasis in relation to PPAR-alpha, PPAR-gamma, and LPL mRNA expressions.  Life Sci. 2005;  77 3344-3354
• 30 Yun SN, Moon SJ, Ko SK, Im BO, Chung SH. Wild ginseng prevents the onset of high-fat diet induced hyperglycemia and obesity in ICR mice.  Arch Pharm Res. 2004;  27 790-796
• 31 Hwang JT, Kim SH, Lee MS, Yang HJ, Kim MJ, Kim HS, Ha J, Kim MS, Kwon DY. Anti-obesity effects of ginsenoside Rh2 are associated with the activation of AMPK signaling pathway in 3T3-L1 adipocyte.  Biochem Biophys Res Commun. 2007;  364 1002-1008

1 These authors contributed equally to this work.

Correspondence