Effects of thyroxine on adhesion molecules and proinflammatory cytokines secretion on human umbilical vein endothelial cells

Attabak Toofani Milani , Mohammad Hassan Khadem-Ansari , Yousef Rasmi


Thyroid dysfunction is associated with elevated cardiovascular risk factors and atherosclerosis. It could be suggested that, hyperthyroidism is related to a higher prevalence of arterial abnormalities. Therefore, evaluating the endothelial dysfunction (ED) related biomarkers seem to be an important issue. It is not clear whether endothelial cells are biologically responsive to thyroid hormones (THs) or how THs induces the production of endothelial cells (EC)-derived proinflammatory mediators. Hence, in this study the effects of thyroxine (T4) on ED and inflammatory related mediators were evaluated. Human umbilical vein endothelial cells was used as endothelial cell model which was treated with concentrations of 50, 100, 200 nmol/L of T4 in various exposure times. In the following, gene and protein expression levels of EC-related markers including intercellular adhesion molecule-1 (ICAM-1), vascular endothelial growth factor (VEGF), and E-selectin were determined using real time polymerase chain reaction (RT-PCR) and western blotting methods. Also, interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) protein levels as proinflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA) method.  Gene and protein expression analysis revealed that T4 treatments up regulated the levels of VEGF, ICAM-1, and E-selectin as ED markers. In addition, T4-treated cells had higher significant levels of IL-6 and TNF-α versus untreated cells in different incubation times. This study proposed the atherosclerotic effects of thyroid hormone. Based on our findings, T4 had strong effects on the gene and protein expression levels of pro-inflammatory, angiogenesis, and ED major mediators associated with atherosclerosis development.


E-selectin; ICAM-1; Interleukin-6; Thyroxine; TNF-α; VEGF.

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Herrington W, Lacey B, Sherliker P, Armitage J, Lewington S. Epidemiology of atherosclerosis and the potential to reduce the global burden of atherothrombotic disease. Circ Res. 2016;118(4):535-546.

Bano A, Chaker L, Mattace-Raso FU, van der Lugt A, Ikram MA, Franco OH, et al. Thyroid function and the risk of atherosclerotic cardiovascular morbidity and mortality: the Rotterdam Study. Circ Res. 2017:CIRCRESAHA. 117.311603.

Burggraaf J, Lalezari S, Emeis J, Vischer U, De Meyer P, Pijl H, et al. Endothelial function in patients with hyperthyroidism before and after treatment with propranolol and thiamazol. Thyroid. 2001;11(2):153-160.

Popławska-Kita A, Siewko K, Telejko B, Modzelewska A, Myśliwiec J, Milewski R, et al. The changes in the endothelial function and haemostatic and inflammatory parameters in subclinical and overt hyperthyroidism. Int J Endocrinol. 2013;2013. DOI: 10.1155/2013/981638.

Coban E, Aydemir M, Yazicioglu G, Ozdogan M. Endothelial dysfunction in subjects with subclinical hyperthyroidism. J Endocrinol Invest. 2006;29(3): 197-200.

Jabbar A, Pingitore A, Pearce SH, Zaman A, Iervasi G, Razvi S. Thyroid hormones and cardiovascular disease. Nat Rev Cardiol. 2017;14(1):39-55.

Naya M, Tsukamoto T, Morita K, Katoh C, Furumoto T, Fujii S, et al. Plasma interleukin-6 and tumor necrosis factor-α can predict coronary endothelial dysfunction in hypertensive patients. Hypertens Res. 2007;30(6):541-548.

Ridker PM, Rifai N, Stampfer MJ, Hennekens CH. Plasma concentration of interleukin-6 and the risk of future myocardial infarction among apparently healthy men. Circulation. 2000;101(15):1767-1772.

Wassmann S, Stumpf M, Strehlow K, Schmid A, Schieffer B, Böhm M, et al. Interleukin-6 induces oxidative stress and endothelial dysfunction by overexpression of the angiotensin II type 1 receptor. Circ Res. 2004;94(4):534-541.

Ridker PM, Rifai N, Pfeffer M, Sacks F, Lepage S, Braunwald E. Elevation of tumor necrosis factor-α and increased risk of recurrent coronary events after myocardial infarction. Circulation. 2000;101(18): 2149-2153.

Esteve E, Castro A, López-Bermejo A, Vendrell J, Ricart W, Fernández-Real JM. Serum interleukin-6 correlates with endothelial dysfunction in healthy men independently of insulin sensitivity. Diabetes Care. 2007;30(4):939-945.

Zhang H, Park Y, Wu J, ping Chen X, Lee S, Yang J, et al. Role of TNF-α in vascular dysfunction. Clin Sci. 2009;116(3):219-230.

Liao JK. Linking endothelial dysfunction with endothelial cell activation. J Clin Invest. 2013;123(2):540-541.

Yuan J, Guo Q, Qureshi AR, Anderstam B, Eriksson M, Heimbürger O, et al. Circulating vascular endothelial growth factor (VEGF) and its soluble receptor 1 (sVEGFR-1) are associated with inflammation and mortality in incident dialysis patients. Nephrol Dial Transplant. 2013;28(9): 2356-2363.

Wang J, Milosveski V, Schramek C, Fong G, Becks G, Hill D. Presence and possible role of vascular endothelial growth factor in thyroid cell growth and function. J Endocrinol. 1998;157(1):5-12.

Davis PJ, Davis FB, Mousa SA. Thyroid hormone-induced angiogenesis. Curr Cardiol Rev. 2009;5(1):12-16.

Liu X, Zheng N, Shi Y-N, Yuan J, Li L. Thyroid hormone induced angiogenesis through the integrin αvβ3/protein kinase D/histone deacetylase 5 signaling pathway. J Mol Endocrinol. 2014;52(3):245-254.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. methods. 2001;25(4): 402-408.

Clynes M, editor. Animal Cell Culture Techniques. Springer Berlin Heidelberg; 2012. P. 54-67.

Kruger NJ. The Bradford Method for Protein Quantitation. In: Walker JM, editor.The Protein Protocols Handbook. Humana Press; 2002. pp. 15-21.

Schägger H, Von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987;166(2):368-379.

Cabral MD, Teixeira P, Soares D, Leite S, Salles E, Waisman M. Effects of thyroxine replacement on endothelial function and carotid artery intima-media thickness in female patients with mild subclinical hypothyroidism. Clinics. 2011;66(8):1321-1328.

Dietrich J, Kuchler-Bopp S, Boutillier S, Ittel M, Reeber A, Zaepfel M, et al. Expression of thyroid hormone receptors alpha and beta-1 messenger RNAs in human endothelial cells. The T3 hormone stimulates the synthesis of the messenger RNA of the intercellular adhesion molecule-1. Cell Mol Biol (Noisy-le-grand). 1997;43(8):1205-1212.

Baumgartner-Parzer S, Wagner L, Reining G, Sexl V, Nowotny P, Müller M, et al. Increase by tri-iodothyronine of endothelin-1, fibronectin and von Willebrand factor in cultured endothelial cells. J Endocrinol. 1997;154(2):231-239.

Fernandez V, Videla LA, Tapia G, Israel Y. Increases in tumor necrosis factor-α in response to thyroid hormone-induced liver oxidative stress in the rat. Free Radic Res. 2002;36(7):719-725.

Lawson C, Wolf S. ICAM-1 signaling in endothelial cells. Pharmacol Rep. 2009;61(1):22-32.

HARA H, SUGITA E, SATO R, BAN Y. Plasma selectin levels in patients with Graves' disease. Endocr J. 1996;43(6):709-713.

Jurgilewicz DH, Rogowski F, Łebkowska U, Citko A, Jaroszewicz E, Parfieńczyk A. E-selectin, L-selectin, ICAM-1 and IL-6 concentrations changes in the serum of patients with hyperthyroidism in the early period of radioiodine I-131 therapy. Nucl Med Rev. 2002;5(1):39-42.

Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA. Vascular endothelial growth factor and angiogenesis. Pharmacol Rev. 2004;56(4):549-580.

Milani AT, Khadem-Ansari MH, Rasmi Y. Effects of thyroid-stimulating hormone on adhesion molecules and pro-inflammatory cytokines secretion in human umbilical vein endothelial cells. Res Pharm Sci. 2018;13(6):546-556.

Szmitko PE, Wang C-H, Weisel RD, de Almeida JR, Anderson TJ, Verma S. New markers of inflammation and endothelial cell activation: Part I. Circulation. 2003;108(16):1917-1923.


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