Organ preservation solutions attenuate accumulation and nuclear translocation of hypoxia-inducible factor-1α in the hepatoma cell line HepG2

Hypoxia-inducible factor-1 alpha (HIF-1 alpha) is a key transcription factor orchestrating hypoxic and inflammatory reactions Here. we determined the impact of organ preservation solutions (Celsior: histidine-tryptophan-ketoglutarate Solution. HTK: University of Wisconsin solution: UW), oxygen supply, and temperature oil HIF-1 alpha accumulation. recorded by Western blotting and immunocytochemistry. in the human hepatoma cell line HepG2 Generation of reactive oxygen species (ROS), NO. and cell viability were concomitantly assessed, At 4 C, HIF-1 alpha accumulation was not detectable in normothermic (37 C) cell culture medium (Dulbecco's; Modified Eagle's Medium, DMEM). HepG2 cells accumulated HIF-1 alpha even in normoxia (21 %. O) which wits not observed in either of the preservation solutions. This correlated to high generation of NO, a normoxic stabilizer of HIF-1 alpha, and L-arginine content (substrate of NO synthesis) in DMEM, and low NO production and absence of L-arginine in preservation solutions. In normothermic hypoxia up to 24 h. intracellular HIF-1 alpha accumulated in all conditions. but less in preservation solutions compared to DMEM. The inhibitory effect oil accumulation and nuclear translocation was most prominent for HTK, the only solution containing the activator of HIF-1 alpha degradation, alpha-ketoglutarate. Addition of other Intermediates of the tricarbon acid cycle-succinate. fumarate, malate did not alter HIF-1 alpha accumulation, although succinate exhibited it beneficial effect on cell viability, in cold storage. In conclusion, preservation solutions attenuate accumulation and nuclear translocation of the transcription factor HIF-1 alpha, and this property is seemingly related to their chemical composition L-arginine, alpha-ketoglutarate). Thus, it appears feasible to design preservation Solution specifically to modify HIF-1 alpha accumulation and nuclear translocation. (C) Copyright 2009 John Wiley & Sons, Ltd.