Aortic effects of thyroid hormone in male mice

in Journal of Molecular Endocrinology
Correspondence should be addressed to J Mittag:
Restricted access

It is well established that thyroid hormones are required for cardiovascular functions; however, the molecular mechanisms remain incompletely understood, especially the individual contributions of genomic and non-genomic signalling pathways. In this study, we dissected how thyroid hormones modulate aortic contractility. To test the immediate effects of thyroid hormones on vasocontractility, we used a wire myograph to record the contractile response of dissected mouse aortas to the adrenergic agonist phenylephrine in the presence of different doses of T3 (3,3′,5-triiodothyronine). Interestingly, we observed reduced vasoconstriction under low and high T3 concentrations, indicating an inversed U-shaped curve with maximal constrictive capacity at euthyroid conditions. We then tested for possible genomic actions of thyroid hormones on vasocontractility by treating mice for 4 days with 1 mg/L thyroxine in drinking water. The study revealed that in contrast to the non-genomic actions the aortas of these animals were hyperresponsive to the contractile stimulus, an effect not observed in endogenously hyperthyroid TRβ knockout mice. To identify targets of genomic thyroid hormone action, we analysed aortic gene expression by microarray, revealing several altered genes including the well-known thyroid hormone target gene hairless. Taken together, the findings demonstrate that thyroid hormones regulate aortic tone through genomic and non-genomic actions, although genomic actions seem to prevail in vivo. Moreover, we identified several novel thyroid hormone target genes that could provide a better understanding of the molecular changes occurring in the hyperthyroid aorta.


      Society for Endocrinology

Article Information


All Time Past Year Past 30 Days
Abstract Views 81 81 81
Full Text Views 14 14 14
PDF Downloads 2 2 2


Related Articles


  • View in gallery

    Aortic response ex vivo recorded in a wire myograph to phenylephrine (PE) after 5 min prestimulation with solvent (0 nM T3), 1 nM T3 (A), 10 nM T3 (B) or 100 nM T3 (C). Please note that the 1 nM T3 curve serves as euthyroid reference control and is thus included for better comparison in (A, B and C). All values are mean ± s.e.m. of n = 8. ***P < 0.001 for top of curve. (D) Phosphorylation of ERK, AKT and AMPK after prestimulation of aortic rings for 10 min with DMSO, 100 nM T3 or 100 nM rT3 as assessed by Western blot. All values are mean ± s.e.m. of n = 9–10.

  • View in gallery

    (A) Serum total T4 and T3 levels after 4 day treatment with 1 mg/L T4 in drinking water. Values are mean ± s.e.m. of n = 6. **P < 0.01; ***P < 0.001 with unpaired 2-tailed t test. (B) Aortic response ex vivo recorded in a wire myograph to phenylephrine (PE) in mice treated for 4 days with 1 mg/L T4 in drinking water as compared to controls. Values are mean ± s.e.m. of n = 10. *P < 0.05 for top of curve. (C) Aortic response ex vivo recorded in a wire myograph to phenylephrine (PE) in mice lacking TRβ as compared to controls. Values are mean ± s.e.m. of n = 5. *P < 0.05 for bottom of curve. (D) Phosphorylation of ERK, AKT and AMPK of aortic rings from wild type or TRβ knockout mice as assessed by Western blot. All values are mean ± s.e.m. of n = 4.

  • View in gallery

    (A) Overview of the genomic effects of T4 on aortic gene expression in mice treated for 4 days with 1 mg/L T4 in drinking water as compared to controls as studied by micoarray analysis. (B) Weighted gene set enrichment analysis of the microarray results. The numbers on the bars indicate the number of genes found to be significantly changed in the respective pathway. (C) Unbiased hierarchical clustering of the microarray results with the top five candidate genes by P-value in (D), which were validated using qPCR in a different cohort (E). Values are mean ± s.e.m. of n = 4–5. *P < 0.05 with unpaired 2-tailed t test. (F) Correlation of gene expression as assessed by microarray with the individual total T3 and T4 levels of the respective animals of the top correlated genes with P < 0.0001. Tm100, transmembrane protein 100; Penk, preproenkephaline; Hr, hairless; Fbln1, fibulin 1; Elmo1, engulfment and cell motility 1; Pel3, HXXXD-type acyltransferase family protein; Maf, MAF bZIP transcription factor; Pola1, DNA Polymerase Alpha 1; Msantd2, Myb/SANT DNA Binding Domain Containing 2.


ArgravesWSGreeneLMCooleyMAGallagherWM 2003 Fibulins: physiological and disease perspectives. EMBO Reports 4 11271131. (

CaoXKambeFMoellerLCRefetoffSSeoH 2005 Thyroid hormone induces rapid activation of Akt/protein kinase B-mammalian target of rapamycin-p70s6K cascade through phosphatidylinositol 3-kinase in human fibroblasts. Molecular Endocrinology 19 102112. (

Carrillo-SepulvedaMACeravoloGSFortesZBCarvalhoMHTostesRCLaurindoFRWebbRCBarreto-ChavesMLM 2010 Thyroid hormone stimulates NO production via activation of the PI3K/Akt pathway in vascular myocytes. Cardiovascular Research 85 560570. (

Carrillo-SepulvedaMACeravoloGSFurstenauCRMonteiro Pde PdeSBruno-FortesZCarvalhoMHLaurindoFRTostesRCWebbRCBarreto-ChavesML 2013 Emerging role of angiotensin type 2 receptor (AT2R)/Akt/NO pathway in vascular smooth muscle cell in the hyperthyroidism. PLoS ONE 8 e61982. (

ChampionHCBivalacquaTJZadinaJEKastinAJHymanALKadowitzPJ 2002 Role of nitric oxide in mediating vasodilator responses to opioid peptides in the rat. Clinical and Experimental Pharmacology and Physiology 29 229232. (

DelitalaAPOrruMFilighedduFPiliaMGDelitalaGGanauASabaPSDecandiaFScuteriAMarongiuMet al. 2015 Serum free thyroxine levels are positively associated with arterial stiffness in the SardiNIA study. Clinical Endocrinology 82 592597. (

DillmannW 2010 Cardiac hypertrophy and thyroid hormone signaling. Heart Failure Reviews 15 125132. (

EbleJANilandS 2009 The extracellular matrix of blood vessels. Current Pharmaceutical Design 15 13851400. (

FabregatASidiropoulosKGarapatiPGillespieMHausmannKHawRJassalBJupeSKorningerFMcKaySet al. 2016 The reactome pathway knowledge base. Nucleic Acids Research 44 D481D487. (

FlamantFChengSYHollenbergANMoellerLCSamarutJWondisfordFEYenPMRefetoffS 2017 Thyroid hormone signaling pathways: time for a more precise nomenclature. Endocrinology 158 20522057. (

ForrestDErwayLCNgLAltschulerRCurranT 1996a Thyroid hormone receptor beta is essential for development of auditory function. Nature Genetics 13 354357. (

ForrestDHanebuthESmeyneRJEverdsNStewartCLWehnerJMCurranT 1996b Recessive resistance to thyroid hormone in mice lacking thyroid hormone receptor beta: evidence for tissue-specific modulation of receptor function. EMBO Journal 15 30063015. (

GachkarSOelkrugRMartinez-SanchezNRial-PensadoEWarnerAHoefigCSLopezMMittagJ 2017 3-Iodothyronamine induces tail vasodilation Through central action in male mice. Endocrinology 158 19771984. (

GrieveDJFletcherSPitsillidesAABothamKMElliottJ 1999 Effects of oral propylthiouracil treatment on nitric oxide production in rat aorta. British Journal of Pharmacology 127 18. (

HiroiYKimHHYingHFuruyaFHuangZSimonciniTNomaKUekiKNguyenNHScanlanTSet al. 2006 Rapid nongenomic actions of thyroid hormone. PNAS 103 1410414109. (

HondaHIwataTMochizukiTKogoH 2000 Changes in vascular reactivity induced by acute hyperthyroidism in isolated rat aortae. General Pharmacology 34 429434. (

HönesGSRakovHLoganJLiaoXHWerbenkoEPollardASPræstholmSMSiersbækMSRijntjesEGassenJet al. 2017 Noncanonical thyroid hormone signaling mediates cardiometabolic effects in vivo. PNAS 114 E11323E11332. (

IttermannTLorbeerRDorrMSchneiderTQuadratAHesselbarthLWenzelMLehmphulIKohrleJMenselBet al. 2016 High levels of thyroid-stimulating hormone are associated with aortic wall thickness in the general population. European Radiology 26 44904496. (

KalyanaramanHSchwappacherRJoshuaJZhuangSScottBTKlosMCasteelDEFrangosJADillmannWBossGRet al. 2014 Nongenomic thyroid hormone signaling occurs through a plasma membrane-localized receptor. Science Signaling 7 ra48. (

KannistoKRehnmarkSSlatisKWebbPLarssonLGafvelsMEggertsenGPariniP 2014 The thyroid receptor beta modulator GC-1 reduces atherosclerosis in ApoE deficient mice. Atherosclerosis 237 544554. (

KantorovichVPacakK 2010 Pheochromocytoma and paraganglioma. Progress in Brain Research 182 343373. (

KersseboomSvan GuchtALMvan MullemABriganteGFarinaSCarlssonBDonkersJMvan de GraafSFJPeetersRPVisserTJ 2017 Role of the bile acid transporter SLC10A1 in liver targeting of the lipid-lowering thyroid hormone analog eprotirome. Endocrinology 158 33073318. (

KleinIOjamaaK 2001 Thyroid hormone and the cardiovascular system. New England Journal of Medicine 344 501509. (

KonigMZimmerAMSteinerHHolmesPVCrawleyJNBrownsteinMJZimmerA 1996 Pain responses, anxiety and aggression in mice deficient in pre-proenkephalin. Nature 383 535538. (

LuoZFujioYKureishiYRudicRDDaumerieGFultonDSessaWCWalshK 2000 Acute modulation of endothelial Akt/PKB activity alters nitric oxide-dependent vasomotor activity in vivo. Journal of Clinical Investigation 106 493499. (

MartinNPMarron Fernandez de VelascoEMizunoFScappiniELGlossBErxlebenCWilliamsJGStapletonHMGentileSArmstrongDL 2014 A rapid cytoplasmic mechanism for PI3 kinase regulation by the nuclear thyroid hormone receptor, TRbeta, and genetic evidence for its role in the maturation of mouse hippocampal synapses in vivo. Endocrinology 155 37133724. (

MayerlSMullerJBauerRRichertSKassmannCMDarrasVMBuderKBoelenAVisserTJHeuerH 2014 Transporters MCT8 and OATP1C1 maintain murine brain thyroid hormone homeostasis. Journal of Clinical Investigation 124 19871999. (

McAllisterRMGrossenburgVDDelpMDLaughlinMH 1998 Effects of hyperthyroidism on vascular contractile and relaxation responses. American Journal of Physiology 274 E946E953. (

MillerLDMcPhiePSuzukiHKatoYLiuETChengSY 2004 Multi-tissue gene-expression analysis in a mouse model of thyroid hormone resistance. Genome Biology 5 R31. (

MoulakakisKGSokolisDPPerreaDNDosiosTDontasIPoulakouMVDimitriouCASandrisGKarayannacosPE 2007 The mechanical performance and histomorphological structure of the descending aorta in hyperthyroidism. Angiology 58 343352. (

ObuobieKSmithJEvansLMJohnRDaviesJSLazarusJH 2002 Increased central arterial stiffness in hypothyroidism. Journal of Clinical Endocrinology and Metabolism 87 46624666. (

OwenPJYingHLangDTomlinsonDLewisMJChengSYLazarusJH 2007 Endothelial dysfunction in a murine model of thyroid hormone resistance. European Journal of Clinical Investigation 37 390395. (

PantosCITzilalisVGiannakakisSCokkinosDDTzeisSMMalliopoulouVMourouzisIAsimakopoulosPCarageorgiouHVaronosDDet al. 2001 Phenylephrine induced aortic vasoconstriction is attenuated in hyperthyroid rats. International Angiology 20 181186.

PantosCMourouzisCKatramadouMSaranteasTMourouzisIKarageorgiouHTesseromatisCKostopanagiotouGAsimacopoulosPCokkinosDV 2006 Decreased vascular reactivity to alpha1 adrenergic stimulation in the presence of hypothyroid state: a part of an adaptive response? International Angiology 25 216220.

PappasMMourouzisKKarageorgiouHTesseromatisCMourouzisIKostopanagiotouGPantosCCokkinosDV 2009 Thyroid hormone modulates the responsiveness of rat aorta to alpha1-adrenergic stimulation: an effect due to increased activation of beta2-adrenergic signaling. International Angiology 28 474478.

SabioJMRodriguez-MarescaMLunaJDGarcia del RioCVargasF 1994 Vascular reactivity to vasoconstrictors in aorta and renal vasculature of hyperthyroid and hypothyroid rats. Pharmacology 49 257264. (

ThompsonCCBottcherMC 1997 The product of a thyroid hormone-responsive gene interacts with thyroid hormone receptors. PNAS 94 85278532. (

WarnerARahmanASolsjöPGottschlingKDavisBVennstromBArnerAMittagJ 2013 Inappropriate heat dissipation ignites brown fat thermogenesis in mice with a mutant thyroid hormone receptor alpha 1. PNAS 110 1624116246. (

YenPM 2001 Physiological and molecular basis of thyroid hormone action. Physiological Reviews 81 10971142. (

ZhangBKirovSSnoddyJ 2005 WebGestalt: an integrated system for exploring gene sets in various biological contexts. Nucleic Acids Research 33 W741W748. (

ZlohMPerez-DiazNTangLPatelPMackenzieLS 2016 Evidence that diclofenac and celecoxib are thyroid hormone receptor beta antagonists. Life Sciences 146 6672. (


Google Scholar