Cardioprotection of bradykinin at reperfusion involves transactivation of the epidermal growth factor receptor via matrix metalloproteinase-8.
OBJECTIVE
The endogenous autacoid bradykinin (BK) reportedly reduces myocardial infarct size when given exogenously at reperfusion. Muscarinic and opioid G-protein-coupled receptors are equally protective and have been shown to couple through a matrix metalloproteinase (MMP)-dependent transactivation of the epidermal growth factor receptor (EGFR). Here we test whether BK protects the rat heart through the EGFR by an MMP-dependent pathway.


METHODS
Infarct size was measured in isolated perfused rat hearts undergoing 30 min regional ischaemia followed by 120 min reperfusion. In additional studies HL-1 cardiomyocytes were loaded with tetramethylrhodamine ethyl to measure their mitochondrial membrane potential (Psim). Adding the calcium ionophore calcimycin, causes Psim-collapse presumably due to calcium-induced mitochondrial permeability transition.


RESULTS
As expected, BK (100 nmol L(-1)) started 5 min prior to reperfusion reduced infarct size from 38.9 +/- 2.0% of the ischaemic zone in control hearts to 22.2 +/- 3.3% (P < 0.001). Co-infusing the EGFR inhibitor AG1478, the broad-spectrum MMP-inhibitor GM6001, or a highly selective MMP-8 inhibitor abolished BK's protection, thus suggesting an MMP-8-dependent EGFR transactivation in the signalling. Eighty minutes of exposure to calcimycin reduced the mean cell fluorescence to 37.4 +/- 1.8% of untreated cells while BK could partly preserve the fluorescence and, hence, protect the cells (50.5 +/- 2.3%, P < 0.001). The BK-induced mitochondrial protection could again be blocked by AG1478, GM6001 and MMP-8 inhibitor. Finally, Western blotting revealed that BK's protection was correlated with increased phosphorylation of EGFR and its downstream target Akt.


CONCLUSIONS
These results indicate that BK at reperfusion triggers its protective signalling pathway through MMP-8-dependent transactivation of the EGFR.
Methner
C
C
Department of Cardiology, Ernst-Moritz-Arndt University, Greifswald, Germany.
Donat
U
U
Felix
S B
SB
Krieg
T
T
D000818:Animals; D001920:Bradykinin / Q000494:pharmacology / Q000627:therapeutic use; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006321:Heart / Q000187:drug effects*; D007476:Ionophores / Q000494:pharmacology; D020784:Matrix Metalloproteinase 8 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009203:Myocardial Infarction / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D015428:Myocardial Reperfusion Injury / Q000188:drug therapy* / Q000473:pathology; D009206:Myocardium / Q000378:metabolism / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
D004791:Enzyme Inhibitors; D007476:Ionophores; D061965:Matrix Metalloproteinase Inhibitors; D000001:Calcimycin; D066246:ErbB Receptors; D020784:Matrix Metalloproteinase 8; D001920:Bradykinin