Acceleration of hepatocellular energy by idebenone during early reperfusion after cold preservation ameliorates heat shock protein 70 gene expression in a pig liver model.
BACKGROUND
Heat shock proteins (HSPs) are induced in the liver after warm ischemia/reperfusion and are thought to be markers of hepatocellular injury and oxidative stress.


METHODS
The influence of variable periods of cold storage followed by reperfusion on the expression of HSP70 was studied in the isolated perfused pig liver. Organs were harvested and stored in histidine-tryptophan-ketoglutarate solution at 4 degrees C and then perfused (210 min) in a closed water bath (38 degrees C), which subjects the liver to fluctuating outer pressure. The role of energy depletion, reactive oxygen intermediates, Kupffer cells, and circulating leukocytes in HSP70 expression was determined.


RESULTS
HSP70 expression was not detectable in liver tissue before explantation or before reperfusion by Northern blot analysis using a pig HSP70 gene probe. HSP70 expression was observed after reperfusion depending on cold storage time. Kinetics of HSP70 expression monitored by reverse transcriptase polymerase chain reaction showed a rapid increase of mRNA within 1 hr, which was closely associated with delayed recovery of hepatocellular energy charge, as assessed by the ketone body ratio. The inactivation of Kupffer cells, the presence or absence of leukocytes, and the suppression of oxidative stress with the antioxidant idebenone, given during reperfusion, had no influence. However, feeding the animals with idebenone over 7 days before explantation led to a faster recovery of ketone body ratio, paralleled by a substantial suppression of HSP70 expression.


CONCLUSIONS
Our data show that HSP70 expression during reperfusion is mainly dependent on the preceding cold storage time and the consecutive delayed recovery of the hepatocellular energy charge.
Schütz
E
E
Abteilung Klinische Chemie, Georg-August-Universität, Göttingen, Germany.
Wieland
E
E
Heine
L
L
Hensel
A
A
Schmiedl
A
A
Armstrong
V W
VW
Richter
J
J
Schuff-Werner
P
P
Günther
E
E
Oellerich
M
M
D000818:Animals; D001483:Base Sequence; D016227:Benzoquinones / Q000494:pharmacology*; D003080:Cold Temperature; D017124:Conserved Sequence; D017931:DNA Primers; D004734:Energy Metabolism / Q000187:drug effects*; D005260:Female; D005682:Gadolinium / Q000494:pharmacology; D005947:Glucose; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis*; D006982:Hypertonic Solutions; D007511:Ischemia; D007657:Ketone Bodies / Q000378:metabolism; D007700:Kinetics; D007728:Kupffer Cells / Q000187:drug effects / Q000648:ultrastructure; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008353:Mannitol; D009926:Organ Preservation / Q000379:methods*; D016133:Polymerase Chain Reaction; D011189:Potassium Chloride; D011343:Procaine; D012333:RNA, Messenger / Q000096:biosynthesis; D015424:Reperfusion; D013552:Swine; D014158:Transcription, Genetic / Q000187:drug effects*; D014451:Ubiquinone / Q000031:analogs & derivatives
D016227:Benzoquinones; C039984:Bretschneider cardioplegic solution; D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D006982:Hypertonic Solutions; D007657:Ketone Bodies; D012333:RNA, Messenger; D014451:Ubiquinone; D008353:Mannitol; D011343:Procaine; D011189:Potassium Chloride; D005682:Gadolinium; C036619:idebenone; D005947:Glucose; C038958:gadolinium chloride