Abstract
Aim
High glucose-induced oxidative stress has been suggested as one of the mediators of endothelial damage in diabetes. The major endothelial protein, endoglin, has been found overexpressed in the vessels during pathological situations, but little is known about its relation to diabetic vascular complications. To clarify the role of endoglin in endothelial injury, we sought to determine the effects of high and oscillating glucose on its expression.
Materials
Furthermore, the activation of the Krüppel-like factor 6 (KLF-6) and the hypoxia-inducible factor-1α (HIF-1α) as possible regulators of endoglin expression has been evaluated. The possible role of the oxidative stress has been studied evaluating the effects of the antioxidant alpha-lipoic acid (ALA) and the cellular antioxidant response mediated by NAD(P)H:quinine-oxido-reductase-1 (NQO-1) and heme oxygenase-1 (HO-1).
Results
Primary HUVECs were cultured for 21 days in normal, high and oscillating glucose (5, 25 and 5/25 mmol/l every 24 h, respectively) with/without ALA. In oscillating and high glucose total endoglin, its soluble form (sEng), KLF-6 and HIF-1α were significantly increased. Simultaneously, the oxidative DNA stress markers 8-OHdG and H2A.X were elevated. Moreover, ENG gene transcriptional rate increased during glucose exposures concomitantly with increased KLF-6 nuclear translocations. ALA significantly reduced all these phenomena. Interestingly, during oscillating and chronic high glucose, NQO-1 and HO-1 did not increase, but ALA induced their overexpression.
Conclusions
Together, these findings provide novel clue about endoglin in the regulation of high glucose-mediated vascular damage in HUVECs and the role of oxidative stress in this regulation.
Similar content being viewed by others
References
Giacco F, Brownlee M (2010) Oxidative stress and diabetic complications. Circ Res 107:1058–1070
Ceriello A, Testa R (2009) Antioxidant anti-inflammatory treatment in type 2 diabetes. Diabetes Care 32(Suppl 2):S232–S236
Ceriello A (2010) Hyperglycaemia and the vessel wall: the pathophysiological aspects on the atherosclerotic burden in patients with diabetes. Eur J Cardiovasc Prev Rehabil 17(Suppl 1):S15–S19
Quagliaro L, Piconi L, Assaloni R, Martinelli L, Motz E, Ceriello A (2003) Intermittent high glucose enhances apoptosis related to oxidative stress in human umbilical vein endothelial cells: the role of protein kinase C and NAD(P)H-oxidase activation. Diabetes 52:2795–2804
Zhu Y, Sun Y, Xie L, Jin K, Sheibani N, Greenberg DA (2003) Hypoxic induction of endoglin via mitogen-activated protein kinases in mouse brain microvascular endothelial cells. Stroke 34:2483–2488
Bernabeu C, Conley BA, Vary CP (2007) Novel biochemical pathways of endoglin in vascular cell physiology. J Cell Biochem 102:1375–1388
Sánchez-Elsner T, Botella LM, Velasco B, Langa C, Bernabéu C (2002) Endoglin expression is regulated by transcriptional cooperation between the hypoxia and transforming growth factor-beta pathways. J Biol Chem 277:43799–43808
Atkins GB, Jain MK (2007) Role of Krüppel-like transcription factors in endothelial biology. Circ Res 100:1686–1695
Torsney E, Charlton R, Parums D, Collis M, Arthur HM (2002) Inducible expression of human endoglin during inflammation and wound healing in vivo. Inflamm Res 51:464–470
Lopez-Novoa JM, Bernabeu C (2010) The physiological role of endoglin in the cardiovascular system. Am J Physiol Heart Circ Physiol 299:H959–H974
Hawinkels LJ, Kuiper P, Wiercinska E, Verspaget HW, Liu Z, Pardali E, Sier CF, ten Dijke P (2010) Matrix metalloproteinase-14 (MT1-MMP)-mediated endoglin shedding inhibits tumor angiogenesis. Cancer Res 70:4141–4150
Valbuena-Diez AC, Blanco FJ, Oujo B, Langa C, Gonzalez-Nuñez M, Llano E, Pendas AM, Díaz M, Castrillo A, Lopez-Novoa JM, Bernabeu C (2012) Oxysterol-induced soluble endoglin release and its involvement in hypertension. Circulation 126:2612–2624
Kaitu’u-Lino TJ, Palmer KR, Whitehead CL, Williams E, Lappas M, Tong S (2012) MMP-14 is expressed in preeclamptic placentas and mediates release of soluble endoglin. Am J Pathol 180:888–894
Blázquez-Medela AM, García-Ortiz L, Gómez-Marcos MA, Recio-Rodríguez JI, Sánchez-Rodríguez A, López-Novoa JM, Martínez-Salgado C (2010) Increased plasma soluble endoglin levels as an indicator of cardiovascular alterations in hypertensive and diabetic patients. BMC Med 8:86
Qi W, Chen X, Holian J, Tan CY, Kelly DJ, Pollock C (2009) Transcription factors Kruppel-like factor 6 and peroxisome proliferator-activated receptor-{gamma} mediate high glucose-induced thioredoxin-interacting protein. Am J Pathol 175:1858–1867
Botella LM, Sánchez-Elsner T, Sanz-Rodriguez F, Kojima S, Shimada J, Guerrero-Esteo M, Cooreman MP, Ratziu V, Langa C, Vary CP, Ramirez JR, Friedman S, Bernabéu C (2002) Transcriptional activation of endoglin and transforming growth factor-beta signaling components by cooperative interaction between Sp1 and KLF6: their potential role in the response to vascular injury. Blood 100:4001–4010
Cullingford TE, Butler MJ, Marshall AK, el Tham L, Sugden PH, Clerk A (2008) Differential regulation of Krüppel-like factor family transcription factor expression in neonatal rat cardiac myocytes: effects of endothelin-1, oxidative stress and cytokines. Biochim Biophys Acta 178:1229–1236
Stärkel P, Sempoux C, Leclercq I, Herin M, Deby C, Desager JP, Horsmans Y (2003) Oxidative stress, KLF6 and transforming growth factor-beta up-regulation differentiate non-alcoholic steatohepatitis progressing to fibrosis from uncomplicated steatosis in rats. J Hepatol 39:538–546
Holian J, Qi W, Kelly DJ, Zhang Y, Mreich E, Pollock CA, Chen XM (2008) Role of Kruppel-like factor 6 in transforming growth factor-beta1-induced epithelial-mesenchymal transition of proximal tubule cells. Am J Physiol Renal Physiol 295:F1388–F1396
Pan Y, Mansfield KD, Bertozzi CC, Rudenko V, Chan DA, Giaccia AJ, Simon MC (2007) Multiple factors affecting cellular redox status and energy metabolism modulate hypoxia-inducible factor prolyl hydroxylase activity in vivo and in vitro. Mol Cell Biol 27:912–925
Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70:1469–1480
Liu Z, Jia X, Duan Y, Xiao H, Sundqvist KG, Permert J, Wang F (2013) Excess glucose induces hypoxia-inducible factor-1α in pancreatic cancer cells and stimulates glucose metabolism and cell migration. Cancer Biol Ther 14:428–435
Wu LL, Chiou CC, Chang PY, Wu JT (2004) Urinary 8-OHdG: a marker of oxidative stress to DNA and a risk factor for cancer, atherosclerosis and diabetics. Clin Chim Acta 339:1–9
Schisano B, Tripathi G, McGee K, McTernan PG, Ceriello A (2011) Glucose oscillations, more than constant high glucose, induce p53 activation and a metabolic memory in human endothelial cells. Diabetologia 54:1219–1226
Ihnat MA, Thorpe JE, Kamat CD, Szabó C, Green DE, Warnke LA, Lacza Z, Cselenyák A, Ross K, Shakir S, Piconi L, Kaltreider RC, Ceriello A (2007) Reactive oxygen species mediate a cellular ‘memory’ of high glucose stress signalling. Diabetologia 50:1523–1531
Shay KP, Moreau RF, Smith EJ, Smith AR, Hagen TM (2009) Alpha-lipoic acid as a dietary supplement: molecular mechanisms and therapeutic potential. Biochim Biophys Acta 1790:1149–1160
Lebrin F, Goumans MJ, Jonker L, Carvalho RL, Valdimarsdottir G, Thorikay M, Mummery C, Arthur HM, ten Dijke P (2004) Endoglin promotes endothelial cell proliferation and TGF-beta/ALK1 signal transduction. EMBO J 23:4018–4028
Rossi E, Sanz-Rodriguez F, Eleno N, Düwell A, Blanco FJ, Langa C, Botella LM, Cabañas C, Lopez-Novoa JM, Bernabeu C (2013) Endothelial endoglin is involved in inflammation: role in leukocyte adhesion and transmigration. Blood 121:403–415
Abu El-Asrar AM, Nawaz MI, Kangave D, Abouammoh M, Mohammad G (2012) High-mobility group box-1 and endothelial cell angiogenic markers in the vitreous from patients with proliferative diabetic retinopathy. Mediators Inflamm 2012:697489
Docherty NG, López-Novoa JM, Arevalo M, Düwel A, Rodriguez-Peña A, Pérez-Barriocanal F, Bernabeu C, Eleno N (2006) Endoglin regulates renal ischaemia-reperfusion injury. Nephrol Dial Transplant 21:2106–2119
Ma X, Labinaz M, Goldstein J, Miller H, Keon WJ, Letarte M, O’Brien E (2000) Endoglin is overexpressed after arterial injury and is required for transforming growth factor-beta-induced inhibition of smooth muscle cell migration. Arterioscler Thromb Vasc Biol 20:2546–2552
van Laake LW, van den Driesche S, Post S, Feijen A, Jansen MA, Driessens MH, Mager JJ, Snijder RJ, Westermann CJ, Doevendans PA, van Echteld CJ, ten Dijke P, Arthur HM, Goumans MJ, Lebrin F, Mummery CL (2006) Endoglin has a crucial role in blood cell-mediated vascular repair. Circulation 114:2288–2297
Suzuki T, Aizawa K, Matsumura T, Nagai R (2005) Vascular implications of the Krüppel-like family of transcription factors. Arterioscler Thromb Vasc Biol 25:1135–1141
Piconi L, Quagliaro L, Assaloni R, Da Ros R, Maier A, Zuodar G, Ceriello A (2006) Constant and intermittent high glucose enhances endothelial cell apoptosis through mitochondrial superoxide overproduction. Diabetes Metab Res Rev 22:198–203
Quagliaro L, Piconi L, Assaloni R, Da Ros R, Szabó C, Ceriello A (2007) Primary role of superoxide anion generation in the cascade of events leading to endothelial dysfunction and damage in high glucose treated HUVEC. Nutr Metab Cardiovasc Dis 17:257–267
Wu CY, Kang HY, Yang WL, Wu J, Jeong YS, Wang J, Chan CH, Lee SW, Zhang X, Lamothe B, Campos AD, Darnay BG, Lin HK (2011) Critical role of monoubiquitination of histone H2AX protein in histone H2AX phosphorylation and DNA damage response. J Biol Chem 286:30806–30815
Packer L, Witt EH, Tritschler HJ (1995) Alpha-lipoic acid as a biological antioxidant. Free Radical Biol Med 19:227–250
Sun LQ, Chen YY, Wang X, Li XJ, Xue B, Qu L, Zhang TT, Mu YM, Lu JM (2012) The protective effect of alpha lipoic acid on Schwann cells exposed to constant or intermittent high glucose. Biochem Pharmacol 84:961–973
Beiroa D, Romero-Picó A, Langa C, Bernabeu C, López M, López-Novoa JM, Nogueiras R, Diéguez C (2013) Heterozygous deficiency of endoglin decreases insulin and hepatic triglyceride levels during high fat diet. PLoS one 8:e54591
Ungvari Z, Bailey-Downs L, Sosnowska D, Gautam T, Koncz P, Losonczy G, Ballabh P, de Cabo R, Sonntag WE, Csiszar A (2011) Vascular oxidative stress in aging: a homeostatic failure due to dysregulation of NRF2-mediated antioxidant response. Am J Physiol Heart Circ Physiol 301:H363–H372
Wang Y, Ying L, Chen YY, Shen YL, Guo R, Jin KK, Wang LX (2014) Induction of heme oxygenase-1 ameliorates vascular dysfunction in streptozotocin-induced type 2 diabetic rats. Vascul Pharmacol 61:16–24
Cudmore M, Ahmad S, Al-Ani B, Fujisawa T, Coxall H, Chudasama K, Devey LR, Wigmore SJ, Abbas A, Hewett PW, Ahmed A (2007) Negative regulation of soluble Flt-1 and soluble endoglin release by heme oxygenase-1. Circulation 115:1789–1797
Conflict of interest
Lucia La Sala, Gemma Pujadas, Valeria De Nigris, Silvia Canivell, Anna Novials, Stefano Genovese, Antonio Ceriello declare that they have no conflict of interest.
Human and animal rights statement
This article does not contain any studies with human or animal subjects performed by any of the authors.
Author information
Authors and Affiliations
Corresponding author
Additional information
Managed by Massimo Federici.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
La Sala, L., Pujadas, G., De Nigris, V. et al. Oscillating glucose and constant high glucose induce endoglin expression in endothelial cells: the role of oxidative stress. Acta Diabetol 52, 505–512 (2015). https://doi.org/10.1007/s00592-014-0670-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00592-014-0670-3