Down-regulation of cardioprotective bradykinin type-2 receptors in the left ventricle of patients with end-stage heart failure
Antti Kuoppala, MD*,
Naotaka Shiota, MD, PhD*,
Jorma O. Kokkonen, MD, PhD* ,
Inka Liesmaa, MD*,
Karam Kostner, MD, PhD ,
Mikko Mäyränpää, MD*,
Petri T. Kovanen, MD, PhD* and
Ken A. Lindstedt, PhD*,*
* Wihuri Research Institute, Helsinki, Finland
Division of Cardiology, Helsinki University Central Hospital, Helsinki, Finland
Division of Cardiology, Allegemeines Krankenhaus, Vienna, Austria
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Figure 1 Quantitative reverse transcription-polymerase chain reaction of bradykinin type-2 receptors (BK-2Rs) in human hearts. (A) Messenger ribonucleic acid (mRNA) expression of BK-2Rs in normal and failing human hearts. (B) The mRNA levels of BK-2Rs are shown as the log ratios between the target and its competitor in the three patient groups. The solid triangle indicates a patient with coronary heart disease (CHD) who did not receive an angiotensin-converting enzyme inhibitor. (C) Expression levels of angiotensin II type I and II receptors (AT-1R and AT-2R, respectively) were analyzed in normal and failing hearts. (D) Levels of BK-2Rs were determined in bradykinin (BK)- and/or captopril (Capto)-treated cultured human coronary artery endothelial cells (HCAEC). *p = 0.01. **p = 0.003. IDC = idiopathic dilated cardiomyopathy; M = marker.
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Figure 2 Western blotting of bradykinin type-2 receptors (BK-2Rs) in human hearts. (A) Representative Western blot showing the expression of BK-2R proteins in normal and failing human hearts. Equal amounts of protein (30 µg/lane) were loaded onto the gel. (B) The relative amounts of BK-2Rs were quantitated in the different patient groups by measuring the optical densities (OD) in the Western blots. *p = 0.04. **p = 0.002. CHD = coronary heart disease; IDC = idiopathic dilated cardiomyopathy.
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Figure 3 Detection of bradykinin type-2 receptors (BK-2Rs) and fibrosis in human hearts. Serial sections of normal (A and D), idiopathic dilated cardiomyopathy (IDC) (B and E) and coronary heart disease (CHD) (C and F) hearts were analyzed for BK-2R expression (A to C) by using a monoclonal BK-2R antibody (magnification x200) and for the degree of fibrosis (D to F) by Masson trichrome staining (magnification x200).
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Figure 4 Detection of bradykinin type-2 receptors (BK-2Rs) and myocardial cells in human hearts. (A) Serial frozen sections of normal hearts were analyzed for cell-specific BK-2R expression (magnification x400) and for specific antibodies in endothelial cells (EC), smooth muscle cells (SMC) and fibroblasts (Fb). A BK-2R–positive myocyte (upper left panel, long black arrow and inset showing the selected myocyte at a higher magnification [x1,000]), an EC (long white arrow), a SMC (short white arrow) and a Fb (short black arrow) are also indicated in the upper left panel. (B) Frozen sections of normal, idiopathic dilated cardiomyopathy (IDC) and coronary heart disease (CHD) hearts were stained for BK-2R expression (magnification x200).
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Figure 5 Expression of endothelial nitric oxide (eNOS) in normal and failing hearts. (A) Expression of eNOS in normal and failing human hearts was analyzed by Western blotting. Equal amounts of protein (50 µg/lane) were run on the gel, and eNOS was quantitated as described in the Methods section. **p = 0.001 for idiopathic dilated cardiomyopathy (IDC) vs. normal and p = 0.002 for coronary heart disease (CHD) vs. normal. OD = optical density.
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