Download PDF
Thromb Haemost 2007; 97(06): 988-997
DOI: 10.1160/TH06-10-0593
Endothelium and Vascular Development
Schattauer GmbH

Biphasic effect of pioglitazone on isolated human endothelial progenitor cells: Involvement of peroxisome proliferator-activated receptor-γ and transforming growth factor-β1

Santiago Redondo
1   Department of Pharmacology, School of Medicine, Universidad Complutense, Madrid, Spain
2   Institute for Molecular Cardiovascular Research (IMCAR)
,
Mihail Hristov
2   Institute for Molecular Cardiovascular Research (IMCAR)
3   Interdisciplinary Center for Clinical Research “BIOMAT”, RWTH University Hospital, Aachen, Germany
,
Denis Gümbel
2   Institute for Molecular Cardiovascular Research (IMCAR)
,
Teresa Tejerina
1   Department of Pharmacology, School of Medicine, Universidad Complutense, Madrid, Spain
,
Christian Weber
2   Institute for Molecular Cardiovascular Research (IMCAR)
3   Interdisciplinary Center for Clinical Research “BIOMAT”, RWTH University Hospital, Aachen, Germany
› Author Affiliations
Further Information

Publication History

Received 17 October 2006

Accepted after resubmission 06 March 2007

Publication Date:
27 November 2017 (online)

    Permissions and Reprints

Summary

Endothelial progenitor cells (EPCs) have been implicated in vascular repair and found to be functionally impaired in patients with diabetes. We evaluated the effects of the anti-diabetic drug pioglitazone on human EPC function and the involvement of PPAR-γ and TGF-β1. EPCs in culture were characterized at day 7 by the development of colony-forming units (CFUs) and flow cytometry assessment of differentiation marker (DiI-ac-LDL/lectin, KDR and CD31). Adhesion on fibronectin and fibrinogen in flow was analyzed as functional parameter. Treatment with pioglitazone for 72 hours increased the number of EPC-CFUs, DiI-ac-LDL+/lectin+, CD31+ and KDR+ EPCs at 1 μM but not at 10 μM. Since pioglitazone did not significantly alter proliferation and apoptosis in cultured EPCs, the increase in EPC number was most likely attributable to augmented adhesion and differentiation. Indeed, pioglitazone increased EPC adhesion in flow at 1 μM, an effect prevented by PPAR-γ and β2-integrin blockade. In contrast, pioglitazone did not promote EPC adhesion at 10 μM; however, increased adhesion became evident by co-incubation with a blocking TGF-β1 antibody. As determined by ELISA, pioglitazone induced a persistent increase in TGF-β1 secretion only at 10 μM when a significantly elevated expression of endoglin, the accessory receptor forTGF-β1, was also observed. Taken together, pioglitazone exerts biphasic effects on the function of isolated EPCs, causing a PPAR-γ-dependent stimulation at 1 μM and a TGF-β1-mediated suppression at 10 μM. These results may help to define optimal therapeutic doses of pioglitazone for improving endothelial dysfunction.

Keywords

Endothelial - pioglitazone - pharmacology - growth factors - adhesion
 

  • References

  • 1 Hristov M, Weber C. Endothelial progenitor cells: characterization, pathophysiology, and possible clinical relevance. J Cell Mol Med 2004; 8: 498-508.
    CrossrefPubMedGoogle Scholar
  • 2 Rabelink TJ, de Boer HC, de Koning EJP. et al. Endothelial progenitor cells: more than an inflammatory response?. ArteriosclerThrombVasc Biol 2004; 24: 834-838.
    PubMedGoogle Scholar
  • 3 Kalka C, Masuda H, Takahashi T. et al. Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci USA 2000; 97: 3422-3427.
    CrossrefPubMedGoogle Scholar
  • 4 Ceradini DJ, Kulkarni AR, Callaghan MJ. et al. Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1. Nat Med 2004; 10: 858-864.
    CrossrefPubMedGoogle Scholar
  • 5 Hristov M, Zernecke A, Bidzhekov K. et al. Importance of CXC chemokine receptor 2 in the homing of human peripheral blood endothelial progenitor cells to sites of arterial injury. Circ Res 2007; 100: 590-597.
    CrossrefPubMedGoogle Scholar
  • 6 Hill JM, Zalos G, Halcox JP. et al. Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. N Engl J Med 2003; 348: 593-600.
    CrossrefPubMedGoogle Scholar
  • 7 Heiss C, Keymel S, Niesler U. et al. Impaired progenitor cell activity in age-related endothelial dysfunction. J Am Coll Cardiol 2005; 45: 1441-1448.
    CrossrefPubMedGoogle Scholar
  • 8 Leor J, Marber M. Endothelial progenitors: a new Tower of Babel?. JAm Coll Cardiol 2006; 48: 1588-1590.
    PubMedGoogle Scholar
  • 9 Vasa M, Fichtlscherer S, Adler K. et al. Increase in circulating endothelial progenitor cells by statin t apy in patients with stable coronary artery disease. Circulation 2001; 103: 2885-2890.
    CrossrefPubMedGoogle Scholar
  • 10 Hristov M, Fach C, Becker C. et al. Reduced numbers of circulating endothelial progenitor cells in patients with coronary artery disease associated with long-term statin treatment.Atherosclerosis. 2006 Epub ahead of print.
    PubMedGoogle Scholar
  • 11 Vijan S, Hayward RA. Pharmacologic lipid-lowering therapy in type 2 diabetes mellitus: background paper for the American College of Physicians. Ann Intern Med 2004; 140: 650-658.
    CrossrefPubMedGoogle Scholar
  • 12 Segal MS, Shah R, Afzal A. et al. Nitric oxide cytoskeletal- induced alterations reverse the endothelial progenitor cell migratory defect associated with diabetes. Diabetes 2006; 55: 102-109.
    CrossrefPubMedGoogle Scholar
  • 13 Tepper OM, Galiano RD, Capla JM. et al. Human endothelial progenitor cells from type II diabetics exhibit impaired proliferation, adhesion, and incorporation into vascular structures. Circulation 2002; 106: 2781-2786.
    CrossrefPubMedGoogle Scholar
  • 14 Campia U, Matuskey LA, Panza JA. Peroxisome proliferator-activated receptor-gamma activation with pioglitazone improves endothelium-dependent dilation in nondiabetic patients with major cardiovascular risk factors. Circulation 2006; 113: 867-875.
    CrossrefPubMedGoogle Scholar
  • 15 Pfutzner A, Marx N, Lubben G. et al. Improvement of cardiovascular risk markers by pioglitazone is independent from glycemic control: results from the pioneer study. J Am Coll Cardiol 2005; 45: 1925-1931.
    CrossrefPubMedGoogle Scholar
  • 16 Dormandy JA, Charbonnel B, Eckland DJ. et al. PROactive investigators. Secondary prevention of macrovascular events in patients with type 2 diabetes in the PROactive Study (PROspective pioglitAzone Clinical Trial In macroVascular Events): a randomised controlled trial. Lancet 2005; 366: 1279-1289.
    CrossrefPubMedGoogle Scholar
  • 17 Wang CH, Ciliberti N, Li SH. et al. Rosiglitazone facilitates angiogenic progenitor cell differentiation toward endothelial lineage: a new paradigm in glitazone pleiotropy. Circulation 2004; 109: 1392-1300.
    CrossrefPubMedGoogle Scholar
  • 18 Goldberg RB, Kendall DM, Deeg MA. et al. GLAI Study Investigators. A comparison of lipid and glycemic effects of pioglitazone and rosiglitazone in patients with type 2 diabetes and dyslipidemia. Diabetes Care 2005; 28: 1547-1554.
    CrossrefPubMedGoogle Scholar
  • 19 Redondo S, Ruiz E, Santos-Gallego CG. et al. Pioglitazone induces vascular smooth muscle cell apoptosis through a peroxisome proliferator-activated receptorgamma, transforming growth factor-beta1, and a Smad2-dependent mechanism. Diabetes 2005; 54: 811-817.
    CrossrefPubMedGoogle Scholar
  • 20 Henrich D, Hahn P, Wahl M. et al. Serum derived from multiple trauma patients promotes the differentiation of endothelial progenitor cells in vitro: possible role of transforming growth factor-beta1 and vascular endothelial growth factor165. Shock 2004; 21: 13-16.
    PubMedGoogle Scholar
  • 21 Zernecke A, Weber KS, Erwig LP. et al. Combinatorial model of chemokine involvement in glomerular monocyte recruitment: role of CXC chemokine receptor 2 in infiltration during nephrotoxic nephritis. J Immunol 2001; 166: 5755-5762.
    CrossrefPubMedGoogle Scholar
  • 22 Blindt R, Vogt F, Astafieva I. et al. A novel drugeluting stent coated with an integrin-binding cyclic Arg-Gly-Asp peptide inhibits neointimal hyperplasia by recruiting endothelial progenitor cells. J Am Coll Cardiol 2006; 47: 1786-1795.
    CrossrefPubMedGoogle Scholar
  • 23 Lebrin F, Goumans MJ, Jonker L. et al. Endoglin promotes endothelial cell proliferation and TGF-beta/ALK1 signal transduction. EMBO J 2004; 23: 4018-4028.
    CrossrefPubMedGoogle Scholar
  • 24 Marx N, Wohrle J, Nusser T. et al. Pioglitazone reduces neointima volume after coronary stent implantation: a randomized, placebo-controlled, double-blind trial in nondiabetic patients. Circulation 2005; 112: 2792-2798.
    CrossrefPubMedGoogle Scholar
  • 25 Budde K, Neumayer HH, Fritsche L. et al. The pharmacokinetics of pioglitazone in patients with impaired renal function. Br J Clin Pharmacol 2003; 55: 368-374.
    CrossrefPubMedGoogle Scholar
  • 26 Verma S, Kuliszewski MA, Li SH. et al. C-reactive protein attenuates endothelial progenitor cell survival, differentiation, and function: further evidence of a mechanistic link between C-reactive protein and cardiovascular disease. Circulation 2004; 109: 2058-2067.
    CrossrefPubMedGoogle Scholar
  • 27 Pistrosch F, Herbrig K, Oelschlaegel U. et al. PPARgamma-agonist rosiglitazone increases number and migratory activity of cultured endothelial progenitor cells. Atherosclerosis 2005; 183: 163-167.
    CrossrefPubMedGoogle Scholar
  • 28 Staels B, Fruchart JC. Therapeutic roles of peroxisome proliferator-activated receptor agonists. Diabetes 2005; 54: 2460-2470.
    CrossrefPubMedGoogle Scholar
  • 29 Gensch C, Clever YP, Werner C. et al. The PPARgamma agonist pioglitazone increases neoangiogenesis and prevents apoptosis of endothelial progenitor cells.Atherosclerosis. 2006 Epub ahead of print.
    PubMedGoogle Scholar
  • 30 Walter DH, Rittig K, Bahlmann FH. et al. Statin therapy accelerates reendothelialization: a novel effect involving mobilization and incorporation of bone marrow- derived endothelial progenitor cells. Circulation 2002; 105: 3017-3024.
    CrossrefPubMedGoogle Scholar
  • 31 Grip O, Janciauskiene S, Lindgren S. Atorvastatin activates PPAR-gamma and attenuates the inflammatory response in human monocytes. Inflamm Res 2002; 51: 58-62.
    CrossrefPubMedGoogle Scholar
  • 32 Chavakis E, Aicher A, Heeschen C. et al. Role of ß2-integrins for homing and neovascularization capacity of endothelial progenitor cells. J Exp Med 2005; 201: 63-72.
    CrossrefPubMedGoogle Scholar
  • 33 Duan L, Cheng H, Sun X. et al. LFA-1 and VLA-4 involved in human high proliferative potential-en endothelial progenitor cells homing to ischemic tissue. Thromb Haemost 2006; 96: 807-815.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 34 Schaefer K, Wada K, Takahashi H. et al. Peroxisome proliferator-activated receptor gamma inhibition prevents adhesion to the extracellular matrix and induces anoikis in hepatocellular carcinoma cells. Cancer Res 2005; 65: 2251-2259.
    CrossrefPubMedGoogle Scholar
  • 35 Fukunaga Y, Itoh H, Doi K. et al. Thiazolidinediones, peroxisome proliferator-activated receptor gamma agonists, regulate endothelial cell growth and secretion of vasoactive peptides. Atherosclerosis 2001; 158: 113-119.
    CrossrefPubMedGoogle Scholar
  • 36 Sassa Y, Hata Y, Aiello LP. et al. Bifunctional properties of peroxisome proliferator-activated receptor gamma1 in KDR gene regulation mediated via interaction with both Sp1 and Sp3. Diabetes 2004; 53: 1222-1229.
    CrossrefPubMedGoogle Scholar
  • 37 Vogt T, Hafner C, Bross K. et al. Antiangiogenetic therapy with pioglitazone, rofecoxib, and metronomic trofosfamide in patients with advanced malignant vascular tumors. Cancer 2003; 98: 2251-2256.
    CrossrefPubMedGoogle Scholar
  • 38 Panigrahy D, Huang S, Kieran MW. et al. PPARgamma as a therapeutic target for tumor angiogenesis and metastasis. Cancer Biol Ther 2005; 4: 687-693.
    CrossrefPubMedGoogle Scholar
  • 39 Sjolie AK, Moller F. Medical management of diabetic retinopathy. Diabet Med 2004; 21: 666-672.
    CrossrefPubMedGoogle Scholar
  • 40 Sales VL, Engelmayr GC, Mettler BA. et al. Transforming growth factor-beta1 modulates extracellular matrix production, proliferation, and apoptosis of endothelial progenitor cells intissue-engineering scaffolds. Circulation 2006; 114: I193-1199.
    PubMedGoogle Scholar
  • 41 Grainger DJ. Transforming growth factor beta and atherosclerosis: so far, so good for the protective cytokine hypothesis. Arterioscler Thromb Vasc Biol 2004; 24: 399-404.
    CrossrefPubMedGoogle Scholar
  • 42 Laviades C, Varo N, Diez J. Transforming growth factor beta in hypertensives with cardiorenal damage. Hypertension 2000; 36: 517-522.
    CrossrefPubMedGoogle Scholar
  • 43 Yamamoto K, Morishita R, Tomita N. et al. Ribozyme oligonucleotides against transforming growth factor-beta inhibited neointimal formation after vascular injury in rat model: potential application of ribozyme strategy to treat cardiovascular disease. Circulation 2000; 102: 1308-1314.
    CrossrefPubMedGoogle Scholar
  • 44 Lutgens E, Gijbels M, Smook M. et al. Transforming growth factor-beta mediates balance between inflammation and fibrosis during plaque progression. Arterioscler Thromb Vasc Biol 2002; 22: 975-982.
    CrossrefPubMedGoogle Scholar
  • 45 Watabe T, Nishihara A, Mishima K. et al. TGF-beta receptor kinase inhibitor enhances growth and integrity of embryonic stem cell-derived endothelial cells. J Cell Biol 2002; 163: 1303-1311.
    PubMedGoogle Scholar