Glutathione peroxidase knockout mice are susceptible to myocardial ischemia reperfusion injury.
BACKGROUND
To test our hypothesis that intracellular antioxidant enzymes constitute a cellular defense against acute stress, we studied myocardial ischemia reperfusion injury in the setting of reduced level of glutathione peroxidase using GSHPx-1 gene knockout mice.


METHODS
Knockout mice were developed by disrupting the coding sequence of GSHPx-1 gene after inserting a neomycin resistance gene derived from pMCIpol A into the EcoRI site located in exon 2. Isolated perfused hearts were prepared from two groups of mice-knockout and nontransgenic controls. A 4-0 silk was attached to the apex of the heart which in turn was attached to a force transducer. Hearts were perfused by the Langendorff mode, and after 20 minutes of stabilization subjected to 30 minutes of ischemia followed by 2 hours of reperfusion. The force developed by the heart (DF) and the first derivative of DF (dF/dt) were recorded. Creatine kinase (CK) release was measured in the perfusate and the infarct size was measured at the end of each experiment.


RESULTS
For both GSHPx-1 knockout and nontransgenic control groups, DF and dF/dt were significantly lower during early postischemic reperfusion compared with baseline, but these values were significantly higher for the control group than the knockout mice throughout most of the reperfusion period. CK release from the heart increased during reperfusion for both groups, but this increase was significantly lower for the control group. The infarct size was also smaller for the control mice as compared with knockouts.


CONCLUSIONS
The results indicate that the knockout mice are more susceptible to ischemia reperfusion injury, suggesting the importance of GSHPx-1 gene in myocardial protection from ischemic reperfusion injury.
Yoshida
T
T
University of Connecticut School of Medicine, Farmington 06030-1110, USA.
Maulik
N
N
Engelman
R M
RM
Ho
Y S
YS
Magnenat
J L
JL
Rousou
J A
JA
Flack
J E
JE
Deaton
D
D
Das
D K
DK
D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000235:genetics / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D009203:Myocardial Infarction / Q000209:etiology; D015428:Myocardial Reperfusion Injury / Q000209:etiology*
D005979:Glutathione Peroxidase; D003402:Creatine Kinase