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Horm Metab Res 2008; 40(11): 760-766
DOI: 10.1055/s-0028-1082041
Original Basic

© Georg Thieme Verlag KG Stuttgart · New York

Variety of Angiotensin Receptors in 3T3-L1 Preadipose Cells and Differentiated Adipocytes

F. Weiland 1 , E. J. Verspohl 1
  • 1Department of Pharmacology, Institute of Pharmaceutical and Medicinal Chemistry, University of Münster, Münster, Germany
Further Information

Publication History

received 20.02.2008

accepted 15.04.2008

Publication Date:
18 August 2008 (online)

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Abstract

A local paracrine acting angiotensin (ANG) system of preadipocytes and mature adipocytes is involved in metabolic effects and tissue differentiation. The present study reports on the investigation of binding affinities for various angiotensin receptors including their relevance in 3T3-L1 adipocytes and preadipocytes and 3T3-442A preadipocytes. Competitive binding studies using both 125I-ANG II and its more stable analogue 125I-SARILE for investigating AT1/AT2 binding sites in 3T3-L1 preadipocytes reveal a biphasic competition curve with KDs at a low and high nanomolar range. By using the AT2 receptor selective ligand 125I-CGP4112A the presence of high affinity AT2 binding sites in preadipocytes was observed. High nonspecific binding and a low receptor number is characteristic for all these experiments. An AT4 binding site (binding site for ANG IV) exists in 3T3-L1 and F442A preadipocytes and adipocytes with a high nanomolar KD. This low binding affinity was confirmed by a biological assay, the IRAP assay (=insulin regulated aminopeptidase assay). IRAP is associated with the AT4 receptor, which is a binding site at the luminal part of membrane bound IRAP. The curves for competition binding and for inhibition of IRAP activity are superimposable with respect to angiotensin IV. In conclusion, AT1 and AT2 binding sites are present in preadipocytes. AT2 receptor binding affinities are shown in preadipocytes for the first time. The description of a non-AT1/AT2 binding site with low affinity remains speculative albeit of high interest because antidiabetic and obesity related effects of angiotensin peptides and sartanes as antagonists are observed at these high concentrations. Local concentrations of ANG II and their degradation products may be extremely high. The low amounts of AT1 and AT2 binding sites emphasize the relevance of other binding sites in adipose tissue development and metabolic effects. The AT4 binding site seems to be one of the predominant receptors in adipose cells. Other degraded, but still bioactive peptides like ANG III, IV and ANG(1–7), activating receptors not influenced by ANG II, could be of importance.

Key words

adipocytes - angiotensin receptors - angiotensin IV - IRAP

References

  • 1 Fischer-Posovsky P, Wabitsch M, Hochberg Z. Endocrinology of adipose tissue – An update.  Horm Metab Res. 2007;  39 314-321
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Frayn KN, Tan GD, Karpe F. Adipose tissue: a key target for diabetes pathophysiology and treatment.  Horm Metab Res. 2007;  39 739-742
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Pfeiffer AFH. Adipose tissue and diabetes therapy: Do we hit the target?.  Horm Metab Res. 2007;  39 734-738
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Karlsson C, Lindell K, Ottosson M, Sjostrom L, Carlsson B, Carlsson LMS. Human adipose tissue expresses angiotensinogen and enzymes required for its conversion to angiotensin II.  J Clin Endocrinol Metab. 1998;  83 3925-3929
    CrossrefPubMedGoogle Scholar
  • 5 Janke J, Engel S, Gorzelniak K, Feldpausch M, Heintze U, Böhnke J, Wellner M, Herse F, Lassalle P, Luft FC, Sharma AM. Adipose tissue and circulating endothelial cell specific molecule-1 in human obesity.  Horm Metab Res. 2006;  38 28-33
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Schling P, Loffler G. Cross talk between adipose tissue cells: impact on pathophysiology.  News Physiol Sci. 2002;  17 99-104
    PubMedGoogle Scholar
  • 7 Schling P, Mallow H, Trindl A, Loffler G. Evidence for a local renin angiotensin system in primary cultured human preadipocytes.  Int J Obes Relat Metab Disord. 1999;  23 336-341
    CrossrefPubMedGoogle Scholar
  • 8 Frayn KN. Adipose tissue as a buffer for daily lipid flux.  Diabetologia. 2002;  45 1201-1210
    CrossrefPubMedGoogle Scholar
  • 9 Engeli S, Bohnke J, Gorzelniak K, Janke J, Schling P, Bader M, Luft FC, Sharma AM. Weight loss and the renin-angiotensin-aldosterone system.  Hypertension. 2005;  45 356-362
    CrossrefPubMedGoogle Scholar
  • 10 Wang C-H, Leung N, Lapointe N, Szeto L, Uffelman KD, Giacca A, Rouleau JL, Lewis GF. Vasopeptidase inhibitor omapatrilat induces profound insulin sensitization and increases myocardial glucose uptake in zucker fatty rats: studies comparing a vasopeptidase inhibitor, angiotensin-converting enzyme inhibitor, and angiotensin II type I receptor blocker.  Circulation. 2003;  107 1923-1929
    CrossrefPubMedGoogle Scholar
  • 11 Henriksen EJ, Jacob S, Kinnick TR, Teachey MK, Krekler M. Selective angiotensin II receptor receptor antagonism reduces insulin resistance in obese Zucker rats.  Hypertension. 2001;  38 884-890
    CrossrefPubMedGoogle Scholar
  • 12 Hansson L, Lindholm LH, Niskanen L, Lanke J, Hedner T, Niklason A, Luomanmaki K, Dahlof B, Faire U de, Morlin C, Karlberg BE, Wester PO, Bjorck JE. Effect of angiotensin-converting-enzyme inhibition compared with conventional therapy on cardiovascular morbidity and mortality in hypertension: the Captopril Prevention Project (CAPPP) randomised trial.  Lancet. 1999;  353 611-616
    CrossrefPubMedGoogle Scholar
  • 13 Chica-Rodriguez S, Cortés-Denia P, Ramirez-Expósito MJ, Martinez-Martos JM. Effects of α1-adrenergic receptor blockade by doxazosin on renin-angiotensin system-regulated aminopeptidase and vasopressin-degrading activities in male and female rat thalamus.  Horm Metab Res. 2007;  39 813-817
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Yvan-Charvet L, Even P, Bloch-Faure M, Guerre-Millo M, Moustaid-Moussa N, Ferre P, Quignard-Boulange A. Deletion of the angiotensin type 2 receptor (AT2R) reduces adipose cell size and protects from diet-induced obesity and insulin resistance.  Diabetes. 2005;  54 991-999
    CrossrefPubMedGoogle Scholar
  • 15 Hsieh P-S, Tai Y-H, Loh C-H, Shih K-C, Cheng W-T, Chu C-H. Functional interaction of AT1 and AT2 receptors in fructose-induced insulin resistance and hypertension in rats.  Metabolism. 2005;  54 157-164
    CrossrefPubMedGoogle Scholar
  • 16 Fujimoto M, Masuzaki H, Tanaka T, Yasue S, Tomita T, Okazawa K, Fujikura J, Chusho H, Ebihara K, Hayashi T, Hosoda K, Nakao K. An angiotensin II AT1 receptor antagonist, telmisartan augments glucose uptake and GLUT4 protein expression in 3T3-L1 adipocytes.  FEBS Lett. 2004;  576 492-497
    CrossrefPubMedGoogle Scholar
  • 17 Catar RA, Müller G, Heidler J, Schmitz G, Bornstein SR, Morawietz . Low-density lipoproteins induce the renin-angiotensin system and their receptors in human endothelial cells.  Horm Metab Res. 2007;  39 801-805
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Gorzelniak K, Engeli S, Janke J, Luft FC, Sharma AM. Hormonal regulation of the human adipose-tissue renin-angiotensin system: relationship to obesity and hypertension.  J Hypertens. 2002;  20 965-973
    CrossrefPubMedGoogle Scholar
  • 19 Mallow H, Trindl A, Loffler G. Production of angiotensin II receptors type one (AT1) and type two (AT2) during the differentiation of 3T3-L1 preadipocytes.  Horm Metab Res. 2000;  32 500-503
    PubMedGoogle Scholar
  • 20 Schling P, Loffler G. Effects of angiotensin II on adipose conversion and expression of genes of the renin-angiotensin system in human preadipocytes.  Horm Metab Res. 2001;  33 189-195
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Jones BH, Standridge MK, Moustaid N. Angiotensin II increases lipogenesis in 3T3-L1 and human adipose cells.  Endocrinology. 1997;  138 1512-1519
    CrossrefPubMedGoogle Scholar
  • 22 Crandall DL, Armellino DC, Busler DE, MacHendry-Rinde B, Kral JG. Angiotensin II receptors in human preadipocytes: role in cell cycle regulation.  Endocrinology. 1999;  140 154-158
    CrossrefPubMedGoogle Scholar
  • 23 Crandall DL, Herzlinger HE, Saunders BD, Armellino DC, Kral JG. Distribution of angiotensin II receptors in rat and human adipocytes.  J Lipid Res. 1994;  35 1378-1385
    PubMedGoogle Scholar
  • 24 Sharma AM, Janke J, Gorzelniak K, Engeli S, Luft FC. Angiotensin blockade prevents type 2 diabetes by formation of fat cells.  Hypertension. 2002;  40 609-611
    CrossrefPubMedGoogle Scholar
  • 25 Janke J, Engeli S, Gorzelniak K, Luft FC, Sharma AM. Mature adipocytes inhibit in vitro differentiation of human preadipocytes via angiotensin type 1 receptors.  Diabetes. 2002;  51 1699-1707
    CrossrefPubMedGoogle Scholar
  • 26 Mendes ACR, Ferreira AJ, Pinheiro SVB, Santos RAS. Chronic infusion of angiotensin-(1-7) reduces heart angiotensin II levels in rats.  Regul Pept. 2005;  125 29-34
    CrossrefPubMedGoogle Scholar
  • 27 Schupp M, Janke J, Clasen R, Unger T, Kintscher U. Angiotensin type 1 receptor blockers induce peroxisome proliferator-activated receptor-gamma activity.  Circulation. 2004;  109 2054-2057
    CrossrefPubMedGoogle Scholar
  • 28 Santos RA, Campagnole-Santos MJ, Andrade SP. Angiotensin-(1-7): an update.  Regul Pept. 2000;  91 45-62
    CrossrefPubMedGoogle Scholar
  • 29 Santos RA, Simoes e Silva AC, Maric C, Silva DM, Machado RP, Buhr I de, Heringer-Walther S, Pinheiro SV, Lopes MT, Bader M, Mendes EP, Lemos VS, Campagnole-Santos MJ, Schultheiss HP, Speth R, Walther T. Angiotensin-(1-7) is an endogenous ligand for the G protein-coupled receptor Mas.  Proc Natl Acad Sci USA. 2003;  100 8258-8263
    CrossrefPubMedGoogle Scholar
  • 30 Hall KL, Hanesworth JM, Ball AE, Felgenhauer GP, Hosick HL, Harding JW. Identification and characterization of a novel angiotensin binding site in cultured vascular smooth muscle cells that is specific for the hexapeptide (3-8) fragment of angiotensin II, angiotensin IV.  Regul Pept. 1993;  44 225-232
    CrossrefPubMedGoogle Scholar
  • 31 Albiston AL, MacDowall SG, Matsacos D, Sim P, Clune E, Mustafa T, Lee J, Mendelsohn FAO, Simpson RJ, Connolly LM, Chai SY. Evidence that the angiotensin IV (AT4) receptor is the enzyme insulin-regulated aminopeptidase.  J Biol Chem. 2001;  276 48623-48626
    CrossrefPubMedGoogle Scholar
  • 32 Schling P. Expression of angiotensin II receptors type 1 and type 2 in human preadipose cells during differentiation.  Horm Metab Res. 2002;  34 709-715
    Article in Thieme ConnectPubMedGoogle Scholar
  • 33 Lew RA, Mustafa T, Ye S, MacDowall SG, Chai SY, Albiston AL. Angiotensin AT4 ligands are potent, competitive inhibitors of insulin regulated aminopeptidase (IRAP).  J Neurochem. 2003;  86 344-350
    CrossrefPubMedGoogle Scholar
  • 34 Herbst JJ, Ross SA, Scott HM, Bobin SA, Morris NJ, Lienhard GE, Keller SR. Insulin stimulates cell surface aminopeptidase activity toward vasopressin in adipocytes.  Am J Physiol Endocrinol Metab. 1997;  272 E600-E606
    PubMedGoogle Scholar
  • 35 AbdAlla S, Lother H, Abdel-tawab AM, Quitterer U. The angiotensin II AT2 receptor is an AT1 receptor antagonist.  J Biol Chem. 2001;  276 39721-39726
    CrossrefPubMedGoogle Scholar
  • 36 Juan C-C, Chien Y, Wu L-Y, Yang W-M, Chang C-L, Lai Y-H, Ho P-H, Kwok CF, Ho L-T. Angiotensin II enhances insulin sensitivity in vitro and in vivo.  Endocrinology. 2005;  2004-1136
    PubMedGoogle Scholar
  • 37 Fliser D, Arnold U, Kohl B, Hartung R, Ritz E. Angiotensin II enhances insulin sensitivity in healthy volunteers under euglycemic conditions.  J Hypertens. 1993;  11 983-988
    CrossrefPubMedGoogle Scholar
  • 38 Caron AZ, Arguin G, Guillemette G. Angiotensin IV interacts with a juxtamembrane site on AT(4)/IRAP suggesting an allosteric mechanism of enzyme modulation.  Regul Pept. 2003;  113 9-15
    CrossrefPubMedGoogle Scholar
  • 39 Keller SR, Scott HM, Mastick CC, Aebersold R, Lienhard GE. Cloning and characterization of a novel insulin-regulated membrane aminopeptidase from Glut4 vesicles.  J Biol Chem. 1995;  270 23612-23618
    CrossrefPubMedGoogle Scholar
  • 40 Keller SR, Davis AC, Clairmont KB. Mice deficient in the insulin-regulated membrane aminopeptidase show substantial decreases in glucose transporter GLUT4 levels but maintain normal glucose homeostasis.  J Biol Chem. 2002;  277 17677-17686
    CrossrefPubMedGoogle Scholar
  • 41 Green H, Meuth M. An established pre-adipose cell line and its differentiation in culture.  Cell. 1974;  3 127-133
    CrossrefPubMedGoogle Scholar
  • 42 Inagami T. Molecular biology and signaling of angiotensin receptors: an overview.  J Am Soc Nephrol. 1999;  10 ((Suppl 11)) S2-S7
    PubMedGoogle Scholar
  • 43 Horiuchi M, Lehtonen JYA, Daviet L. Signaling mechanism of the AT2 angiotensin II receptor: crosstalk between AT1 and AT2 receptors in cell growth.  Trends Endocrinol Metabol. 1999;  10 391-396
    PubMedGoogle Scholar
  • 44 Thomas WG, Mendelsohn FA. Angiotensin receptors: form and function and distribution.  Int J Biochem Cell Biol. 2003;  35 774-779
    CrossrefPubMedGoogle Scholar
  • 45 Gasparo M de, Catt KJ, Inagami T, Wright JW, Unger T. International Union of Pharmacology. XXIII. The Angiotensin II Receptors.  Pharmacol Rev. 2000;  52 415-472
    PubMedGoogle Scholar
  • 46 Zhao Y, Foryst-Ludwig A, Bruemmer D, Culman J, Bader M, Unger T, Kintscher U. Angiotensin II induces peroxisome proliferator-activated receptor gamma in PC12W cells via angiotensin type 2 receptor activation.  J Neurochem. 2005;  94 1395-1401
    CrossrefPubMedGoogle Scholar
  • 47 Clasen R, Schupp M, Foryst-Ludwig A, Sprang C, Clemenz M, Krikov M, Thone-Reineke C, Unger T, Kintscher U. PPAR{gamma}-activating angiotensin type-1 receptor blockers induce adiponectin.  Hypertension. 2005;  46 137-143
    CrossrefPubMedGoogle Scholar
  • 48 MacMullen JR, Gibson KJ, Lumbers ER, Burrell JH. 125I[Sar1Ile8] Angiotensin II has a different affinity for AT1 and AT2 receptor subtypes in ovine tissues.  Regul Pept. 2002;  105 83-92
    CrossrefPubMedGoogle Scholar
  • 49 Greenland K, Wyse B, Sernia C. Identification and characterization of angiotensinIV binding sites in rat neurone and astrocyte cell cultures.  J Neuroendocrinol. 1996;  8 687-693
    CrossrefPubMedGoogle Scholar
  • 50 Ross SA, Keller SR, Lienhard GE. Increased intracellular sequestration of the insulin-regulated aminopeptidase upon differentiation of 3T3-L1 cells.  Biochem J. 1998;  330 1003-1008
    PubMedGoogle Scholar

Correspondence

E. J. Verspohl

Department of Pharmacology

Institute of Pharmaceutical and Medicinal Chemistry

Hittorfstr. 58–62

48149 Münster

Germany

Phone: +49/251/833 33 39

Fax: +49/251/833 21 44

Email: verspoh@uni-muenster.de