Importance of endothelial nitric oxide synthase for the hypothermic protection of lungs against ischemia-reperfusion injury.
OBJECTIVE
The hypothesis that the protective effects of mild hypothermia against the pulmonary ischemia-reperfusion injury are mediated by endothelial nitric oxide synthase was tested.


METHODS
Endothelial nitric oxide synthase knock-out and wild-type mice were sham operated or underwent a 1-hour occlusion of the left pulmonary hilum, followed by 5 hours of reperfusion. Temperature in the left pleural cavity during ischemia was maintained at either 36 degrees C (normothermia) or 32 degrees C (hypothermia). Inflammatory response (myeloperoxidase activity), endothelial barrier function (extravasation of Evans blue-labeled albumin), and endothelial nitric oxide synthase expression and phosphorylation were determined at the end of reperfusion.


RESULTS
After normothermic ischemia both strains had a similar mortality (wild-type, 22.9%; knock-out, 15.4%), which was completely abolished by hypothermia. Endothelial barrier function was disturbed after normothermic ischemia in both wild-type and knock-out mice. Mild hypothermia significantly reduced pulmonary Evans blue extravasation in wild-type mice, but not in knock-out mice. Myeloperoxidase activity increased after normothermic ischemia to the same degree in both strains. This response was significantly attenuated by hypothermia in wild-type mice, but not in knock-out mice. In wild-type mice, endothelial nitric oxide synthase expression and phosphorylation were higher after hypothermic ischemia than after normothermic ischemia. No effect of ischemia on expression of inducible nitric oxide synthase was found in wild-type or knock-out mice.


CONCLUSIONS
Hypothermic protection against pulmonary ischemia-reperfusion injury is dependent on endothelial nitric oxide synthase and is associated with increased expression and phosphorylation of endothelial nitric oxide synthase.
Zhang
Li
L
Department of Cardiac Surgery, University of Rostock, Rostock, Germany.
Kumar
Sanjeev
S
Kaminski
Alexander
A
Kasch
Cornelius
C
Sponholz
Christoph
C
Stamm
Christof
C
Ladilov
Yury
Y
Steinhoff
Gustav
G