Histidine-induced injury to cultured liver cells, effects of histidine derivatives and of iron chelators.
The amino acid histidine is an excellent buffer and is therefore included in several organ preservation solutions used in transplantation medicine. However, when used at concentrations as in these solutions, histidine has a marked injurious potential. Therefore, we here assessed the mechanism of histidine-induced cell injury and searched for ways to use the buffering power of histidine but avoid histidine toxicity. When cultured hepatocytes were incubated in HTK solution or in modified Krebs-Henseleit buffer containing 198 mM L-histidine at 37 degrees C, most cells lost viability within 3 h (LDH release 86 +/- 7% and 89 +/- 5%, respectively). This injury was accompanied by marked lipid peroxidation, and was strongly inhibited by hypoxia, by the antioxidants trolox, butylated hydroxytoluene and N-acetylcysteine and by the membrane-permeable iron chelators 2,2'-dipyridyl, 1,10-phenanthroline, LK 614, LK 616 and deferoxamine. Thus, histidine-induced cell injury appears to be mediated by an iron-dependent formation of reactive oxygen species. D-Histidine, imidazol and L-histidine methyl ester also elicited marked injury, while the N-substituted derivatives Nalpha-acetyl-L-histidine and tert-butyl-oxycarbonylhistidine and histidine-containing dipeptides showed almost no toxicity. Histidine toxicity, its iron dependence and the superiority of Nalpha-acetyl-L-histidine were also evident during/after cold (4 degrees C) incubations. Therefore, we suggest the addition of iron chelators to histidine-containing solutions, and/or replacing histidine with Nalpha-acetyl-L-histidine in organ preservation solutions.
Rauen
U
U
Institut für Physiologische Chemie, Universitätsklinikum, Hufelandstr. 55, 45122, Essen, Germany. ursula.rauen@uni-duisburg-essen.de
Klempt
S
S
de Groot
H
H
D000111:Acetylcysteine / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002084:Butylated Hydroxytoluene / Q000378:metabolism / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D002839:Chromans / Q000378:metabolism / Q000494:pharmacology; D003676:Deferoxamine / Q000378:metabolism / Q000494:pharmacology; D022781:Hepatocytes / Q000187:drug effects*; D006639:Histidine / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000633:toxicity*; D007502:Iron Chelating Agents / Q000378:metabolism* / Q000494:pharmacology; D015227:Lipid Peroxidation / Q000187:drug effects; D008297:Male; D015394:Molecular Structure; D019188:Organ Preservation Solutions / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar
D002839:Chromans; D007502:Iron Chelating Agents; D019188:Organ Preservation Solutions; D002084:Butylated Hydroxytoluene; D006639:Histidine; D003676:Deferoxamine; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D000111:Acetylcysteine