Induction of aquaporin-1 mRNA following cardiopulmonary bypass and reperfusion.
BACKGROUND
Cardiopulmonary bypass (CPB) and hypothermic circulatory arrest (HCA) are important components of congenital cardiac surgery. Ischemia/reperfusion injury and inflammatory cascade activation result in endothelial damage and vascular leak which are clinically manifested as pulmonary edema and low cardiac output postoperatively. Newborns are particularly susceptible. Subtraction cloning is a useful method of isolating induced genes and can be applied to CPB/HCA.


METHODS
We used a newborn lamb model replicating infant CPB with HCA to obtain tissues during various periods of reperfusion. We utilized subtraction cloning to identify mRNA induced in lung following CPB/HCA and reperfusion. Ribonuclease protection was used to quantify mRNA levels.


RESULTS
We isolated a cDNA encoding ovine aquaporin-1 in a subtracted cDNA screen comparing control lung with lung exposed to CPB/HCA and reperfusion. Aquaporin-1 mRNA levels increased 3-fold in lung (p = .006) exposed to CPB/HCA and 6 hr of reperfusion. No induction was observed immediately following bypass or after 3 hr of reperfusion. We found no significant induction of aquaporin-1 mRNA following bypass, arrest, and reperfusion in other tissues surveyed, including ventricle, atrium, skeletal muscle, kidney, brain, and liver.


CONCLUSIONS
Our finding that aquaporin-1 mRNA is reproducibly induced in lung following CPB/HCA with 6 hr of reperfusion suggests an important role for the water channel in the setting of pulmonary edema. Induction of Aquaporin-1 is late compared with other inflammatory mediators (ICAM-1, E-selectin, IL-8). Further studies are needed to determine if aquaporin-1 contributes to the disease process or if it is part of the recovery phase.
Tabbutt
S
S
Department of Cardiology, Children's Hospital, Dana Farber Cancer Institute, Boston.
Nelson
D P
DP
Tsai
N
N
Miura
T
T
Hickey
P R
PR
Mayer
J E
JE
Neufeld
E J
EJ
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D051398:Aquaporin 1; D020346:Aquaporins; D002315:Cardiopulmonary Bypass; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D006324:Heart Arrest, Induced; D006439:Hemodynamics; D007473:Ion Channels / Q000235:genetics*; D008168:Lung / Q000378:metabolism*; D008969:Molecular Sequence Data; D015428:Myocardial Reperfusion Injury / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012756:Sheep; D014867:Water / Q000378:metabolism
D020346:Aquaporins; D018076:DNA, Complementary; D007473:Ion Channels; D012333:RNA, Messenger; D014867:Water; D051398:Aquaporin 1