title: Hypothermia causes a marked injury to rat proximal tubular cells that is aggravated by all currently used preservation solutions.
abstract: Cold preservation results in cell death via iron-dependent formation of reactive oxygen species, leading to apoptosis during rewarming. We aimed to study cold-induced damage (i.e., injury as a consequence of hypothermia itself and not cold ischemia) in proximal tubular cells (PTC) in various preservation solutions presently applied and to clarify the role of mitochondria in this injury. Primary cultures of rat PTC were incubated at 4 degrees C for 24 h in culture medium, UW, Euro-Collins or HTK solution with and without the iron chelator desferal and rewarmed at 37 degrees C in culture medium. Cell damage, morphology, and apoptosis were studied and mitochondrial membrane potential was assessed by fluorescence microscopy. Cold incubation of PTC in culture medium followed by rewarming caused marked cell damage compared to warm incubation alone (LDH release 39+/-10% vs. 1.6+/-0.3%). Cold-induced damage was aggravated in all preservation solutions (LDH release 85+/-2% for UW; similar in Euro-Collins and HTK). After rewarming, cells showed features suggestive for apoptosis. Desferal prevented cell injury in all solutions (e.g., 8+/-2% for UW). Mitochondrial membrane potential was lost during rewarming and this loss could also be inhibited by desferal. Trifluoperazine, which is known to inhibit mitochondrial permeability transition (MPT), was able to prevent cold-induced injury (LDH 85+/-5% vs. 12+/-2%). We conclude that cold-induced injury occurs in PTC and is aggravated by UW, Euro-Collins, and HTK solution. Iron-dependent MPT is suggested to play a role in this damage. Strategies to prevent cold-induced injury should aim at reducing the availability of "free" iron.
authors.affiliation: Department of Pharmacology and Toxicology, University Medical Center Nijmegen, Nijmegen, The Netherlands.
authors.forename: Miranda
authors.initials: M
authors.lastname: Bartels-Stringer
authors.forename: Cornelis
authors.initials: C
authors.lastname: Kramers
authors.forename: Jack F M
authors.initials: JF
authors.lastname: Wetzels
authors.forename: Frans G M
authors.initials: FG
authors.lastname: Russel
authors.forename: Herbert de
authors.initials: Hd
authors.lastname: Groot
authors.forename: Ursula
authors.initials: U
authors.lastname: Rauen
mesh_terms: D000241:Adenosine / Q000494:pharmacology*; D000493:Allopurinol / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D003470:Culture Media / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D005978:Glutathione / Q000494:pharmacology*; D006982:Hypertonic Solutions / Q000494:pharmacology; D007036:Hypothermia, Induced; D007328:Insulin / Q000494:pharmacology*; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000378:metabolism; D008353:Mannitol / Q000494:pharmacology; D008928:Mitochondria / Q000378:metabolism; D019188:Organ Preservation Solutions / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D011189:Potassium Chloride / Q000494:pharmacology; D011343:Procaine / Q000494:pharmacology; D011887:Raffinose / Q000494:pharmacology*; D051381:Rats
chemical_list: C039984:Bretschneider cardioplegic solution; D003470:Culture Media; C037096:Euro-Collins' solution; D006982:Hypertonic Solutions; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D008353:Mannitol; D011343:Procaine; D000493:Allopurinol; D011189:Potassium Chloride; D005978:Glutathione; D005947:Glucose; D000241:Adenosine; D011887:Raffinose