A Phosphodiesterase-5 Inhibitor Vardenafil Enhances Angiogenesis Through a Protein Kinase G-Dependent Hypoxia-Inducible Factor-1/Vascular Endothelial Growth Factor Pathway

Objective—We examined whether phosphodiesterase-5 (PDE5) inhibition can promote ischemia-induced angiogenesis. Methods and Results—Unilateral hindlimb ischemia was generated by resecting right femoral artery in wild-type C3H/He mice, treated with either vehicle or a PDE5 inhibitor vardenafil (10 mg/kg per day). Four weeks after surgery, vardenafil significantly enhanced blood flow recovery and augmented capillary collateral formation in ischemic muscle (blood flow ratios of ischemic/nonischemic leg: 0.52±0.17 [vehicle] versus 0.92±0.09 [vardenafil], P<0.01). Vardenafil upregulated protein expression of vascular endothelial growth factor and hypoxia-inducible factor (HIF)-1&agr; in ischemic muscle and enhanced mobilization of Sca-1/Flk-1-positive endothelial progenitor cells (EPCs) in peripheral blood and bone marrow, contributing to neovascularization. Vardenafil also promoted capillary-like tube formation of human umbilical vein endothelial cells and increased the number of human blood mononuclear cell-derived EPCs in vitro. Furthermore, reporter assays showed that vardenafil and cGMP activated the transactivation activity of HIF-1 under hypoxia. These effects of vardenafil were markedly inhibited by genetic ablation of endothelial nitric oxide synthase, a soluble guanylate cyclase inhibitor, and a protein kinase G inhibitor, respectively. Conclusion—Our results suggest that PDE5 inhibition enhances ischemia-induced angiogenesis with mobilization of EPCs through a protein kinase G-dependent HIF-1/vascular endothelial growth factor pathway. PDE5 inhibition may have a therapeutic potential to treat ischemic cardiovascular diseases.

[1]  Kazuto Nakamura,et al.  Diverse Contribution of Bone Marrow-Derived Cells to Vascular Remodeling Associated With Pulmonary Arterial Hypertension and Arterial Neointimal Formation , 2007, Circulation.

[2]  J. Folkman.  Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.

[3]  Michael I. Wilson,et al.  C. elegans EGL-9 and Mammalian Homologs Define a Family of Dioxygenases that Regulate HIF by Prolyl Hydroxylation , 2001, Cell.

[4]  H. Nishimatsu,et al.  Endothelial nitric oxide synthase is essential for the HMG‐CoA reductase inhibitor cerivastatin to promote collateral growth in response to ischemia , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  R. Kloner,et al.  Therapeutic potential of phosphodiesterase 5 inhibition for cardiovascular disease. , 2003, Circulation.

[6]  G. Semenza,et al.  Nitric Oxide Induces Hypoxia-inducible Factor 1 Activation That Is Dependent on MAPK and Phosphatidylinositol 3-Kinase Signaling* , 2004, Journal of Biological Chemistry.

[7]  D. Lefer,et al.  Sildenafil Promotes Ischemia-Induced Angiogenesis Through a PKG-Dependent Pathway , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[8]  J. Prchal,et al.  Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals. , 2007, Blood.

[9]  M. Endres,et al.  Physical training increases endothelial progenitor cells, inhibits neointima formation, and enhances angiogenesis. , 2003, Circulation.

[10]  G. Semenza,et al.  Regulation of mammalian O2 homeostasis by hypoxia-inducible factor 1. , 1999, Annual review of cell and developmental biology.

[11]  C. Heeschen,et al.  Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells , 2003, Nature Medicine.

[12]  Christie M. Orschell,et al.  Peripheral Blood “Endothelial Progenitor Cells” Are Derived From Monocyte/Macrophages and Secrete Angiogenic Growth Factors , 2003, Circulation.

[13]  G. Palù,et al.  PDE-5 inhibitor, Vardenafil, increases circulating progenitor cells in humans , 2005, International Journal of Impotence Research.

[14]  A. Quyyumi,et al.  Circulating endothelial progenitor cells, vascular function, and cardiovascular risk. , 2003, The New England journal of medicine.

[15]  R Bicknell,et al.  Nitric oxide synthase lies downstream from vascular endothelial growth factor-induced but not basic fibroblast growth factor-induced angiogenesis. , 1997, The Journal of clinical investigation.

[16]  G. Semenza,et al.  Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1 , 1996, Molecular and cellular biology.

[17]  T. Wieland,et al.  The natriuretic peptide/guanylyl cyclase--a system functions as a stress-responsive regulator of angiogenesis in mice. , 2009, The Journal of clinical investigation.

[18]  W. Sessa,et al.  The phosphodiesterase 5 inhibitor sildenafil stimulates angiogenesis through a protein kinase G/MAPK pathway , 2007, Journal of cellular physiology.

[19]  J. Isner,et al.  VEGF contributes to postnatal neovascularization by mobilizing bone marrow‐derived endothelial progenitor cells , 1999, The EMBO journal.

[20]  Jieli Chen,et al.  Nitric Oxide Enhances Angiogenesis via the Synthesis of Vascular Endothelial Growth Factor and cGMP After Stroke in the Rat , 2003, Circulation research.

[21]  J. Beavo,et al.  Regulation of Nitric Oxide–Sensitive Guanylyl Cyclase Cyclic GMP Phosphodiesterases and Regulation of Smooth Muscle Function Structure, Regulation, and Function of Membrane Guanylyl Cyclase Receptors, With a Focus on GC-A Cyclic GMP–Dependent Protein Kinases and the Cardiovascular System: Insights F , 2003 .

[22]  K. Pollok,et al.  Identification of a novel hierarchy of endothelial progenitor cells using human peripheral and umbilical cord blood. , 2004, Blood.

[23]  M. Gassmann,et al.  Oxygen(es) and the hypoxia-inducible factor-1. , 1997, Biological chemistry.

[24]  T F Lue,et al.  Oral sildenafil in the treatment of erectile dysfunction. Sildenafil Study Group. , 1998, The New England journal of medicine.

[25]  Takayuki Asahara,et al.  Isolation of Putative Progenitor Endothelial Cells for Angiogenesis , 1997, Science.

[26]  Martin R. Wilkins,et al.  Phosphodiesterase Type 5 as a Target for the Treatment of Hypoxia-Induced Pulmonary Hypertension , 2003, Circulation.

[27]  Denys N. Wheatley,et al.  An in vitro model of angiogenesis: Basic features , 2004, Angiogenesis.

[28]  J. Corbin,et al.  Tissue distribution of phosphodiesterase families and the effects of sildenafil on tissue cyclic nucleotides, platelet function, and the contractile responses of trabeculae carneae and aortic rings in vitro. , 1999, The American journal of cardiology.

[29]  I. Goldstein,et al.  Oral sildenafil in the treatment of erectile dysfunction. Sildenafil Study Group. , 1998, The New England journal of medicine.

[30]  W. Risau,et al.  Mechanisms of angiogenesis , 1997, Nature.

[31]  S. Dimmeler,et al.  Endothelial Progenitor Cells: Characterization and Role in Vascular Biology , 2004, Circulation research.

[32]  M. Sata.  Role of Circulating Vascular Progenitors in Angiogenesis, Vascular Healing, and Pulmonary Hypertension: Lessons From Animal Models , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[33]  D. Bagger-sjöbäck,et al.  An In Vitro Model , 1987 .