Preservation solutions alter Mrp2-dependent bile flow in cold ischemic rat livers.
BACKGROUND
Previously, decreased organic anion transport through multidrug resistance protein 2 (Mrp2) was observed without any notable cell lysis, even when the livers were stored for 8 hours in University of Wisconsin solution (UW). The aim of this study was to examine the bile flow and its constituents, markers of graft dysfunction without necrosis in cold ischemic livers, using the following preservation media: UW, ET-Kyoto solution (ET-K) and histidine-tryptophan-ketoglutarate solution (HTK).


METHODS
Rat livers were stored at 4 degrees C for 8 hours in the preservation media, and reperfused to collect the bile and determine their constituents. Glycyrrhizin (GL) and/or glutathione (GSH) were added to the media as necessary. The transport efficiency of Mrp2 was assessed by the biliary excretion of 5-carboxyfluorescein (CF), a fluoroprobe excreted from Mrp2. The Intracellular distribution of Mrp2 was determined by immunostaining.


RESULTS
Livers stored for 8 hours exhibited significantly decreased bile production and biliary glutathione (GSH) levels without notable cell lysis. CF excretion was significantly delayed in all solutions. However, these markers were remarkably improved by the redistribution of Mrp2 from the cytoplasm to the canalicular membrane, when the livers were exposed to UW in the presence of GL. Moreover, livers exposed to the Kyoto and HTK solutions increased their bile production and organic anion transport in the presence of GL and GSH.


CONCLUSIONS
These results suggest that the addition of GL and GSH to preservation solutions improves bile production and biliary organic anion transport by increasing Mrp2 localization to the bile canaliculi in post-cold ischemic livers. (248 words).
Sui
Shaoguang
S
Department of Hepatobiliary-Pancreatic Surgery, Graduate School of Medicine, Tokyo Medical and Dental University, Tokyo, Japan.
Kudo
Atsushi
A
Suematsu
Makoto
M
Tanaka
Shinji
S
Nakamura
Noriaki
N
Ito
Koji
K
Arii
Shigeki
S
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D000893:Anti-Inflammatory Agents; D001646:Bile / Q000378:metabolism*; D050377:Cold Ischemia; D005978:Glutathione; D019695:Glycyrrhizic Acid; D008099:Liver / Q000378:metabolism*; D008297:Male; D009926:Organ Preservation; D019188:Organ Preservation Solutions; D051381:Rats; D017208:Rats, Wistar
D018528:ATP-Binding Cassette Transporters; C423013:Abcc2 protein, rat; D000893:Anti-Inflammatory Agents; D019188:Organ Preservation Solutions; D019695:Glycyrrhizic Acid; D005978:Glutathione