Effects of duration of ischemia and donor pretreatment with methylprednisolone or its macromolecular prodrug on the disposition of indocyanine green in cold-preserved rat livers.
OBJECTIVE
Cold preservation of the liver before transplantation may change uptake and excretory functions of hepatocytes. We hypothesized that an increase in the duration of preservation would result in a progressive decrease in the hepatic uptake and/or biliary excretion of indocyanine green (ICG), which would be attenuated by pharmacologic interventions.


METHODS
Donor rats (n = 40) were administered saline (control) or single 5 mg/kg doses of methylprednisolone (MP) or its liver-targeted prodrug (DMP) 2 h prior to liver harvest. Following preservation in cold University of Wisconsin solution for 0, 24, 48, or 72 h, livers were reperfused in a single-pass manner for 30 min in the presence of ICG (approximately 4 microg/ml), followed by 60 min of ICG-free perfusion. The inlet, outlet, and bile concentrations of ICG were measured periodically by high performance liquid chromatography (HPLC), and kinetic parameters were estimated.


RESULTS
Effects of duration of preservation: In unpreserved livers, a significant portion of ICG dose (16%) was effluxed from the liver during the washout period. Cold preservation for 24-72 h progressively increased (p < 0.05) the efflux of ICG (>2-fold at 72 h). Similarly, average extraction ratio showed a modest (30-40%) decrease with increasing preservation time (p < 0.05). However, biliary excretion of ICG showed the most sensitivity to the preservation time (14 to >800-fold decline). Effects of pretreatment: DMP caused significant (p < 0.05) increases in biliary ICG levels (>12-fold) and bile flow rates (6-15-fold) of preserved livers. Although MP pretreatment significantly (p < 0.05) increased (6-fold) bile flow rates in 48-h preserved livers, its effects on biliary ICG levels were not significant (p > 0.05).


CONCLUSIONS
Biliary excretion of ICG is the most sensitive kinetic parameter to prolonged cold ischemia-reperfusion injury in a rat liver perfusion model. The injury may be significantly attenuated by pharmacologic pretreatment of the liver donors.
Chimalakonda
Anjaneya P
AP
School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, Texas 79106, USA.
Mehvar
Reza
R
D000135:Acid Phosphatase / Q000737:chemistry / Q000378:metabolism; D000410:Alanine Transaminase / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000737:chemistry / Q000378:metabolism; D001646:Bile / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D015925:Cryopreservation; D003911:Dextrans / Q000008:administration & dosage / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D004338:Drug Combinations; D007208:Indocyanine Green / Q000378:metabolism* / Q000494:pharmacology; D007511:Ischemia; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D017093:Liver Failure; D016031:Liver Transplantation / Q000379:methods; D008776:Methylprednisolone Hemisuccinate / Q000008:administration & dosage / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D009926:Organ Preservation / Q000379:methods; D019188:Organ Preservation Solutions; D010477:Perfusion / Q000379:methods; D011355:Prodrugs / Q000008:administration & dosage / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury; D013997:Time Factors; D014019:Tissue Donors
D003911:Dextrans; D004338:Drug Combinations; D019188:Organ Preservation Solutions; D011355:Prodrugs; D008776:Methylprednisolone Hemisuccinate; D007770:L-Lactate Dehydrogenase; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase; D000135:Acid Phosphatase; D007208:Indocyanine Green