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Increased Vascular Endothelial Growth Factor Expression But Impaired Vascular Endothelial Growth Factor Receptor Signaling in the Myocardium of Type 2 Diabetic Patients With Chronic Coronary Heart Disease

Ferdinando Carlo Sasso, MD, PhD^_*,,_*, Daniele Torella, MD^,, Ornella Carbonara, MD^_*,, Georgina M. Ellison, PhD^||, Michele Torella, MD^,, Michelangelo Scardone, MD^,, Claudio Marra, MD^,, Rodolfo Nasti, MD^_*,, Raffaele Marfella, MD, PhD^_*,, Domenico Cozzolino, MD^_*,, Ciro Indolfi, MD^,, Maurizio Cotrufo, MD^,, Roberto Torella, MD^_*, and Teresa Salvatore, MD^_*,

^_* Department of Geriatrics and Metabolic Disease, Second University of Naples, Naples, Italy
Department of Cardio-Thoracic and Respiratory Sciences, Second University of Naples, Naples, Italy
Excellence Centre for Cardiovascular Disease, Second University of Naples, Naples, Italy
Division of Cardiology, Magna Graecia University, Catanzaro, Italy
^|| The Research Institute for Sport & Exercise Sciences, Liverpool John Moores University, Liverpool, United Kingdom

View larger version (25K): [in a new window]   Figure 1 (A) Representative Western blot showing vascular endothelial growth factor (VEGF) and VEGF receptors fms-like tyrosine kinase 1 (flt-1) and fetal liver kinase 1 (flk-1) protein expression in the myocardial samples from control human hearts, non-diabetic and diabetic patients with coronary heart disease (CHD). 1 to 3 = control specimens; 4 to 6 = non-diabetic patients with chronic CHD; 7 to 9 = diabetic patients with chronic CHD. (B) Optical density (O.D.) of VEGF, flt-1, and flk-1. *Statistical difference from control specimens; **statistical difference from non-diabetic patients.

View larger version (36K): [in a new window]   Figure 2 (A) Representative reverse transcriptase-polymerase chain reaction (RT-PCR) showing VEGF165 and VEGF receptors (flt-1 and flk-1) messenger ribonucleic acid (mRNA) levels in the myocardial samples from control human hearts, non-diabetic and diabetic patients with CHD. 1 to 3 = control specimens; 4 to 6 = non-diabetic patients with chronic CHD; 7 to 9 = diabetic patients with chronic CHD. (B) Real-time RT-PCR quantification of myocardial VEGF, flt-1 and flk-1 mRNA. Data are the ratio values of VEGF/B-Actin, Flt-1/B-Actin, and Flk-1/B-Actin mRNA (arbitrary units). *Statistical difference from control specimens; **statistical difference from non-diabetic patients. bp = base pair; other abbreviations as in Figure 1.

View larger version (27K): [in a new window]   Figure 3 (A) Representative Western blot showing phospho–flk-1 and Akt and endothelial nitric oxide synthase (eNOS) protein expression and their phosphorylation forms in the myocardial samples from the three groups included in the study. 1 to 3 = control specimens; 4 to 6 = non-diabetic patients with chronic CHD; 7 to 9 = diabetic patients with chronic CHD. (B) Optical density (O.D.) of phospho–flk-1, phospho-Akt, total and phospho-eNOS, and phospho-eNOS/total eNOS ratio. *Statistical difference from control specimens; **statistical difference from non-diabetic patients. Other abbreviations as in Figure 1.