Improved cold preservation of kidney tubular cells by means of adding bioflavonoids to organ preservation solutions.
BACKGROUND
Cold ischemia and reperfusion during renal transplantation result in release of reactive oxygen species. The aim of this study is to examine whether cold storage induced cell injury can be ameliorated by adding flavonoids directly to preservation solutions.


METHODS
Cultured renal tubular epithelial cells (LLC-PK1) were stored in University of Wisconsin (UW) or Euro-Collins (EC) solution at 4 degrees C for 20 hours. Preservation solutions were supplemented with various flavonoids. After rewarming, structural and metabolic cell integrity was measured by lactate dehydrogenase (LDH) release and MTT-test, and lipid peroxidation was assessed from generation of thiobarbituric acid-reactive substances (TBARS).


RESULTS
Twenty hours of cold storage resulted in a substantial loss of cell viability in both preservation solutions (in EC: LDH release 92.4+/-2.7%; MTT-test 0.5+/-0.7%). Addition of luteolin, quercetin, kempferol, fisetin, myricetin, morin, catechin, and silibinin significantly reduced cell injury (for luteolin in EC: LDH release 2.4+/-1.6%; MTT-test 110.3+/-10.4%, P<0.01; TBARS-production (related to cold stored control cells) 8.9+/-2.6%). No cytoprotection was found for apigenin, naringenin, and rutin. Protective potency of flavonoids depends on number of hydroxyl-substituents and lipophilicity of the diphenylpyran compounds.


CONCLUSIONS
Cold storage induced injury of renal tubular cells was substantially ameliorated by adding selected flavonoids directly to preservation solutions.
Ahlenstiel
Thurid
T
Department of Medicine, Division of Nephrology and Hypertension, University Hospital of Saarland, Homburg/Saar, Germany.
Burkhardt
Gunther
G
Köhler
Hans
H
Kuhlmann
Martin K
MK
D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D003080:Cold Temperature; D003451:Cryoprotective Agents / Q000008:administration & dosage; D005419:Flavonoids / Q000008:administration & dosage / Q000737:chemistry; D005978:Glutathione; D006982:Hypertonic Solutions; D066298:In Vitro Techniques; D007328:Insulin; D016030:Kidney Transplantation; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018374:LLC-PK1 Cells; D015227:Lipid Peroxidation / Q000187:drug effects; D047311:Luteolin / Q000008:administration & dosage; D015394:Molecular Structure; D009926:Organ Preservation / Q000379:methods*; D019188:Organ Preservation Solutions; D011794:Quercetin / Q000008:administration & dosage; D011887:Raffinose; D015427:Reperfusion Injury / Q000517:prevention & control; D013552:Swine
D003451:Cryoprotective Agents; C037096:Euro-Collins' solution; D005419:Flavonoids; D006982:Hypertonic Solutions; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D011794:Quercetin; D007770:L-Lactate Dehydrogenase; D005978:Glutathione; D000241:Adenosine; D047311:Luteolin; D011887:Raffinose