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Horm Metab Res 2003; 35(3): 147-152
DOI: 10.1055/s-2003-39075
Original Basic
© Georg Thieme Verlag Stuttgart · New York

Interleukin (IL)-6 mRNA Expression is Stimulated by Insulin, Isoproterenol, Tumour Necrosis Factor Alpha, Growth Hormone, and IL-6 in 3T3-L1 Adipocytes

M.  Fasshauer 1 , J.  Klein 2 , U.  Lossner 1 , R.  Paschke 1
  • 1 University of Leipzig, Department of Internal Medicine III, Leipzig, Germany
  • 2 University of Lübeck, Department of Internal Medicine I, Lübeck, Germany
Further Information

Publication History

Received 26 September 2002

Accepted after revision 5 November 2002

Publication Date:
07 May 2003 (online)

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Abstract

Interleukin (IL)-6 has recently been shown to be an adipocyte-expressed cytokine. Its serum concentrations are elevated in insulin resistance and obesity. For further evaluation of IL-6 gene expression regulation, fully differentiated 3T3-L1 adipocytes were treated with various hormones known to induce insulin resistance. IL-6 mRNA content was measured by quantitative real-time reverse transcription-polymerase chain reaction. Interestingly, treatment of adipocytes with 100 nM insulin, 10 µM isoproterenol, 10 ng/ml tumour necrosis factor alpha (TNFα), and 500 ng/ml growth hormone (GH) for 16 h stimulated IL-6 mRNA expression 2.3-fold, 47-fold, 74-fold, and 1.4-fold, respectively (p < 0.01). In contrast, treatment with 100 nM dexamethasone significantly decreased IL-6 expression to 32 % of control levels (p < 0.01), whereas triiodothyronine and angiotensin 2 did not have any effect. Furthermore, stimulation of IL-6 expression was time-dependent with maximal stimulatory effects detectable after 1 h of insulin, isoproterenol, and GH addition and 12 h of TNFα, respectively. Moreover, isoproterenol's effect could be almost completely reversed by pretreatment of 3T3-L1 cells with the β-adrenergic antagonist propranolol and mimicked by stimulation of GS-proteins with cholera toxin and adenylyl cyclase with forskolin and dibutyryl cAMP, respectively. Finally, IL-6 strongly induced its own expression in a time-dependent fashion. Taken together, our results demonstrate that IL-6 expression in adipocytes is governed by an autocrine positive feedback loop and upregulated by insulin, isoproterenol, TNFα, and GH. In concert with this adipocytokine's upregulation in states of decreased insulin sensitivity such as obesity and diabetes, the data support a possible role of IL-6 as a selectively regulated mediator of insulin resistance.

Key words

3T3-L1 Adipocyte - Growth Hormone - Insulin Resistance - Insulin - Isoproterenol - Obesity - IL-6 - TNFα

References

  • 1 Matthaei S, Stumvoll M, Kellerer M, Haring H U. Pathophysiology and pharmacological treatment of insulin resistance.  Endocr Rev. 2000;  21 585-618
    CrossrefPubMedGoogle Scholar
  • 2 Saltiel A R. The molecular and physiological basis of insulin resistance: emerging implications for metabolic and cardiovascular diseases.  J Clin Invest. 2000;  106 163-164
    CrossrefPubMedGoogle Scholar
  • 3 Spiegelman B M, Flier J S. Obesity and the regulation of energy balance.  Cell. 2001;  104 531-543
    CrossrefPubMedGoogle Scholar
  • 4 Steppan C M, Bailey S T, Bhat S, Brown E J, Banerjee R R, Wright C M, Patel H R, Ahima R S, Lazar M A. The hormone resistin links obesity to diabetes.  Nature. 2001;  409 307-312
    CrossrefPubMedGoogle Scholar
  • 5 Folli F, Kahn C R, Hansen H, Bouchie J L, Feener E P. Angiotensin II inhibits insulin signaling in aortic smooth muscle cells at multiple levels. A potential role for serine phosphorylation in insulin/angiotensin II crosstalk.  J Clin Invest. 1997;  100 2158-2169
    CrossrefPubMedGoogle Scholar
  • 6 Yamauchi T, Kamon J, Waki H, Terauchi Y, Kubota N, Hara K, Mori Y, Ide T, Murakami K, Tsuboyama-Kasaoka N, Ezaki O, Akanuma Y, Gavrilova O, Vinson C, Reitman M L, Kagechika H, Shudo K, Yoda M, Nakano Y, Tobe K, Nagai R, 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
    CrossrefPubMedGoogle Scholar
  • 7 Berg A H, Combs T P, Du X, Brownlee M, Scherer P E. The adipocyte-secreted protein Acrp30 enhances hepatic insulin action.  Nat Med. 2001;  7 947-953
    CrossrefPubMedGoogle Scholar
  • 8 Mohamed-Ali V, Goodrick S, Rawesh A, Katz D R, Miles J M, Yudkin J S, Klein S, Coppack S W. Subcutaneous adipose tissue releases interleukin-6, but not tumor necrosis factor-alpha, in vivo.  J Clin Endocrinol Metab. 1997;  82 4196-4200
    CrossrefPubMedGoogle Scholar
  • 9 Fried S K, Bunkin D A, Greenberg A S. Omental and subcutaneous adipose tissues of obese subjects release interleukin-6: depot difference and regulation by glucocorticoid.  J Clin Endocrinol Metab. 1998;  83 847-850
    CrossrefPubMedGoogle Scholar
  • 10 Pickup J C, Mattock M B, Chusney G D, Burt D. NIDDM as a disease of the innate immune system: association of acute-phase reactants and interleukin-6 with metabolic syndrome X.  Diabetologia. 1997;  40 1286-1292
    CrossrefPubMedGoogle Scholar
  • 11 Vozarova B, Weyer C, Hanson K, Tataranni P A, Bogardus C, Pratley R E. Circulating interleukin-6 in relation to adiposity, insulin action, and insulin secretion.  Obes Res. 2001;  9 414-417
    CrossrefPubMedGoogle Scholar
  • 12 Pradhan A D, Manson J E, Rifai N, Buring J E, Ridker P M. C-reactive protein, interleukin 6, and risk of developing type 2 diabetes mellitus.  JAMA. 2001;  286 327-334
    CrossrefPubMedGoogle Scholar
  • 13 Stith R D, Luo J. Endocrine and carbohydrate responses to interleukin-6 in vivo.  Circ Shock. 1994;  44 210-215
    PubMedGoogle Scholar
  • 14 Tsigos C, Papanicolaou D A, Kyrou I, Defensor R, Mitsiadis C S, Chrousos G P. Dose-dependent effects of recombinant human interleukin-6 on glucose regulation.  J Clin Endocrinol Metab. 1997;  82 4167-4170
    CrossrefPubMedGoogle Scholar
  • 15 Fasshauer M, Klein J, Neumann S, Eszlinger M, Paschke R. Isoproterenol inhibits resistin gene expression through a G(S)-protein-coupled pathway in 3T3-L1 adipocytes.  FEBS Lett. 2001;  500 60-63
    CrossrefPubMedGoogle Scholar
  • 16 Fasshauer M, Klein J, Neumann S, Eszlinger M, Paschke R. Tumor necrosis factor alpha is a negative regulator of resistin gene expression and secretion in 3T3-L1 adipocytes.  Biochem Biophys Res Commun. 2001;  288 1027-1031
    CrossrefPubMedGoogle Scholar
  • 17 Lin J, Puigserver P, Donovan J, Tarr P, Spiegelman B M. Peroxisome proliferator-activated receptor gamma coactivator 1beta (PGC-1beta ), a novel PGC-1-related transcription coactivator associated with host cell factor.  J Biol Chem. 2002;  277 1645-1648
    CrossrefPubMedGoogle Scholar
  • 18 Pickup J C, Crook M A. Is type II diabetes mellitus a disease of the innate immune system?.  Diabetologia. 1998;  41 1241-1248
    CrossrefPubMedGoogle Scholar
  • 19 Kahn B B, Flier J S. Obesity and insulin resistance.  J Clin Invest. 2000;  106 473-481
    CrossrefPubMedGoogle Scholar
  • 20 Bruun J M, Pedersen S B, Richelsen B. Interleukin-8 production in human adipose tissue. inhibitory effects of anti-diabetic compounds, the thiazolidinedione ciglitazone and the biguanide metformin.  Horm Metab Res. 2000;  32 537-541
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Bluher M, Michael M D, Peroni O D, Ueki K, Carter N, Kahn B B, Kahn C R. Adipose tissue selective insulin receptor knockout protects against obesity and obesity-related glucose intolerance.  Dev Cell. 2002;  3 25-38
    CrossrefPubMedGoogle Scholar
  • 22 Guerra C, Navarro P, Valverde A M, Arribas M, Bruning J, Kozak L P, Kahn C R, Benito M. Brown adipose tissue-specific insulin receptor knockout shows diabetic phenotype without insulin resistance.  J Clin Invest. 2001;  108 1205-1213
    CrossrefPubMedGoogle Scholar
  • 23 Rotter V, Nagaev I, Smith U. Interleukin-6 (IL-6) Reduces Gene and Protein Expression of IRS-1 and GLUT4 and Is overexpressed in Human Fat cells from Insulin-Resistant Subjects.  Diabetes (Supplement 2). 2002;  51 A303
    PubMedGoogle Scholar
  • 24 Senn J J, Klover P J, Nowak I, Mooney R A. Interleukin-6 Inhibits Insulin Receptor Signal Transduction in Hepatocytes.  Diabetes (Supplement 2). 2002;  51 A303
    PubMedGoogle Scholar
  • 25 Febbraio M A, Pedersen B K. Muscle-derived interleukin-6: mechanisms for activation and possible biological roles.  FASEB J. 2002;  16 1335-1347
    CrossrefPubMedGoogle Scholar
  • 26 Stouthard J M, Oude E R, Sauerwein H P. Interleukin-6 enhances glucose transport in 3T3-L1 adipocytes.  Biochem Biophys Res Commun. 1996;  220 241-245
    CrossrefPubMedGoogle Scholar
  • 27 Reaven G M, Lithell H, Landsberg L. Hypertension and associated metabolic abnormalities-the role of insulin resistance and the sympathoadrenal system.  N Engl J Med. 1996;  334 374-381
    CrossrefPubMedGoogle Scholar
  • 28 Lawrence V J, Coppack S W. The endocrine function of the fat cell-regulation by the sympathetic nervous system.  Horm Metab Res. 2000;  32 453-467
    PubMedGoogle Scholar
  • 29 Bluher M, Windgassen M, Paschke R. Improvement of insulin sensitivity after adrenalectomy in patients with pheochromocytoma.  Diabetes Care. 2000;  23 1591-1592
    PubMedGoogle Scholar
  • 30 Klein J, Fasshauer M, Ito M, Lowell B B, Benito M, Kahn C R. beta(3)-adrenergic stimulation differentially inhibits insulin signaling and decreases insulin-induced glucose uptake in brown adipocytes.  J Biol Chem. 1999;  274 34 795-34 802
    PubMedGoogle Scholar
  • 31 Fasshauer M, Klein J, Kriauciunas K M, Ueki K, Benito M, Kahn C R. Essential role of insulin receptor substrate 1 in differentiation of brown adipocytes.  Mol Cell Biol. 2001;  21 319-329
    CrossrefPubMedGoogle Scholar
  • 32 Fasshauer M, Klein J, Ueki K, Kriauciunas K M, Benito M, White M F, Kahn C R. Essential role of insulin receptor substrate-2 in insulin stimulation of Glut4 translocation and glucose uptake in brown adipocytes.  J Biol Chem. 2000;  275 25 494-25 501
    PubMedGoogle Scholar
  • 33 Fasshauer M, Klein J, Neumann S, Eszlinger M, Paschke R. Adiponectin gene expression is inhibited by beta-adrenergic stimulation via protein kinase A in 3T3-L1 adipocytes.  FEBS Lett. 2001;  507 142-146
    CrossrefPubMedGoogle Scholar
  • 34 Collins S, Surwit R S. The beta-adrenergic receptors and the control of adipose tissue metabolism and thermogenesis.  Recent Prog Horm Res. 2001;  56 309-328
    CrossrefPubMedGoogle Scholar
  • 35 Gerich J E. The genetic basis of type 2 diabetes mellitus: impaired insulin secretion versus impaired insulin sensitivity.  Endocr Rev. 1998;  19 491-503
    CrossrefPubMedGoogle Scholar
  • 36 Hotamisligil G S, Shargill N S, Spiegelman B M. Adipose expression of tumor necrosis factor-alpha: direct role in obesity-linked insulin resistance.  Science. 1993;  259 87-91
    CrossrefPubMedGoogle Scholar
  • 37 Bluher M, Kratzsch J, Paschke R. Plasma levels of tumor necrosis factor-alpha, angiotensin II, growth hormone, and IGF-I are not elevated in insulin-resistant obese individuals with impaired glucose tolerance.  Diabetes Care. 2001;  24 328-334
    PubMedGoogle Scholar
  • 38 Hotamisligil G S. The role of TNFalpha and TNF receptors in obesity and insulin resistance.  J Intern Med. 1999;  245 621-625
    CrossrefPubMedGoogle Scholar
  • 39 Fasshauer M, Klein J, Neumann S, Eszlinger M, Paschke R. Hormonal regulation of adiponectin gene expression in 3T3-L1 adipocytes.  Biochem Biophys Res Commun. 2002;  290 1084-1089
    CrossrefPubMedGoogle Scholar
  • 40 Rizza R A, Mandarino L J, Gerich J E. Effects of growth hormone on insulin action in man. Mechanisms of insulin resistance, impaired suppression of glucose production, and impaired stimulation of glucose utilization.  Diabetes. 1982;  31 663-669
    CrossrefPubMedGoogle Scholar
  • 41 Takano A, Haruta T, Iwata M, Usui I, Uno T, Kawahara J, Ueno E, Sasaoka T, Kobayashi M. Growth hormone induces cellular insulin resistance by uncoupling phosphatidylinositol 3-kinase and its downstream signals in 3t3-l1 adipocytes.  Diabetes. 2001;  50 1891-1900
    PubMedGoogle Scholar
  • 42 Andrews R C, Walker B R. Glucocorticoids and insulin resistance: old hormones, new targets.  Clin Sci (Colch ). 1999;  96 513-523
    CrossrefPubMedGoogle Scholar
  • 43 Fickova M, Zorad S, Macho L. The effect of in vivo thyroxine treatment on insulin receptors, glucose transport and GLUT4 in rat adipocytes.  Horm Metab Res. 1997;  29 16-19
    PubMedGoogle Scholar
  • 44 Hardie L J, Guilhot N, Trayhurn P. Regulation of leptin production in cultured mature white adipocytes.  Horm Metab Res. 1996;  28 685-689
    PubMedGoogle Scholar
  • 45 Delporte M L, Funahashi T, Takahashi M, Matsuzawa Y, Brichard S M. Pre- and post-translational negative effect of beta-adrenoceptor agonists on adiponectin secretion: in vitro and in vivo studies.  Biochem J. 2002;  367 677-685
    CrossrefPubMedGoogle Scholar
  • 46 Kappes A, Loffler G. Influences of ionomycin, dibutyryl-cycloAMP and tumour necrosis factor-alpha on intracellular amount and secretion of apM1 in differentiating primary human preadipocytes.  Horm Metab Res. 2000;  32 548-554
    PubMedGoogle Scholar

Prof. Dr. R. Paschke

University of Leipzig · Department of Internal Medicine III

Ph.-Rosenthal-Str. 27 · 04103 Leipzig · Germany

Phone: + 49 (341) 97 13-200

Fax: + 49 (341) 97 13-209

Email: pasr@medizin.uni-leipzig.de