Formation of 4-hydroxy-2-nonenal protein adducts in the ischemic rat heart after transplantation.
BACKGROUND
Free radicals formed during ischemia and reperfusion can lead to lipid peroxidation (LPO) and the formation of 4-hydroxy-2-nonenal (4-HNE), one of the most toxic products of LPO. Using a heterotopic rat heart transplantation model we investigated endogenous 4-HNE formation as a response to cold storage of the transplant and warm blood reperfusion in the recipient.


METHODS
Lewis rat hearts were subjected to 30, 240 or 480 minutes of 4 degrees C cold ischemia in Bretschneider cardioplegic solution without or with transplantation and 240-minute reperfusion in F344 recipients. The amount of 4-HNE modified proteins was quantified in rat heart cryosections with an antibody recognizing cysteine-, histidine- and lysine-4-HNE Michael adducts and image analysis of immunostained tissue.


RESULTS
We detected 4-HNE-modified proteins in ischemic rat hearts after transplantation and reperfusion. In hearts submitted to ischemia only, 4-HNE-protein adducts comprised 0.7 +/- 0.3% (30 minutes), 0.7 +/- 0.4% (240 minutes) and 0.2 +/- 0.1% (480 minutes) (mean +/- SEM) of the tissue area. Transplantation and reperfusion in the recipient significantly increased the amount of protein adducts to 6.8 +/- 2.6% (p = 0.041), 5.2 +/- 1.4% (p = 0.009) and 5.7 +/- 0.9% (p = 0.002) in 30-, 240- and 480-minute ischemic hearts, respectively.


CONCLUSIONS
Under the conditions applied in the present study, cold ischemia for >30 minutes did not significantly alter the amount of 4-HNE protein adducts. However, because after transplantation and reperfusion, 6% of heart tissue consisted of 4-HNE-modified proteins, it can be assumed that this damage negatively affects long-term survival of the transplant.
Renner
André
A
Clinic and Policlinic of Cardiothoracic Surgery, University of Wuerzburg, Wuerzburg, Germany. renner_a@klinik.uni-wuerzburg.de
Sagstetter
Martina R
MR
Harms
Harry
H
Lange
Volkmar
V
Götz
Mario E
ME
Elert
Olaf
O
D000447:Aldehydes / Q000378:metabolism*; D000818:Animals; D004195:Disease Models, Animal; D016027:Heart Transplantation; D007091:Image Processing, Computer-Assisted; D008297:Male; D017202:Myocardial Ischemia / Q000378:metabolism*; D015425:Myocardial Reperfusion; D009926:Organ Preservation; D051381:Rats; D011916:Rats, Inbred F344; D011917:Rats, Inbred Lew; D013696:Temperature; D016042:Transplantation, Heterotopic
D000447:Aldehydes; C027576:4-hydroxy-2-nonenal