title: Effect of glutamine on heat-shock-induced mRNA and stress proteins.
abstract: Our aim was to delineate the effect of glutamine on the level of heat shock-inducible mRNA and synthesis of stress protein(s) in cultured kidney cells. Experiments were carried out using opossum kidney (OK) cells. The induction of HSP70 mRNA as well as the synthesis of 72,73 kDa stress proteins was evaluated in cell monolayers exposed to 45 degrees C for 15 minutes followed by a recovery period at 37 degrees C for 3 hours. Incubations were performed in Krebs buffer supplemented with 0, 2, 5, or 10 mM glutamine. A separate series of experiments was performed in the presence of glutamine metabolites, such as NH4Cl, glutamate, or aspartate. Glutamine without preincubation at 37 degrees C remarkably increased the steady-state level of HSP70 mRNA as well as the production of 72,73 kDa stress proteins in a dose-dependent manner. The production of stress protein(s) in the presence of glutamine was associated with decreased percent LDH efflux, suggesting cytoprotective action of glutamine in cultured kidney cells. However, when OK cells were preincubated for 1 hour at 37 degrees C with 10 mM glutamine, there was an approximately fourfold decline in level of HSP70 mRNA compared with experiments in the presence of 10 mM glutamine without preincubation. In addition, metabolites of glutamine, i.e., ammonia and glutamate decreased the level of heat-inducible HSP70 mRNA. Furthermore, aspartate or NH4Cl had little effect on LDH release compared with heat shock experiments, without addition of amino acids. These observations suggest that metabolites of glutamine may blunt the steady-state level of glutamate or HSP70 mRNA. The decreased level of HSP70 mRNA in the presence of NH4Cl may explain the role of ammonia in renal injury and brain toxicity, as well as glutamate excitotoxicity.
authors.affiliation: Department of Pediatrics, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine 19104.
authors.forename: I
authors.initials: I
authors.lastname: Nissim
authors.forename: B
authors.initials: B
authors.lastname: States
authors.forename: M
authors.initials: M
authors.lastname: Hardy
authors.forename: J
authors.initials: J
authors.lastname: Pleasure
authors.forename: I
authors.initials: I
authors.lastname: Nissim
mesh_terms: D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D001224:Aspartic Acid / Q000494:pharmacology; D015152:Blotting, Northern; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D005971:Glutamates / Q000032:analysis / Q000378:metabolism; D018698:Glutamic Acid; D005973:Glutamine / Q000378:metabolism / Q000494:pharmacology*; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature / Q000009:adverse effects*; D007668:Kidney / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000032:analysis / Q000378:metabolism; D009893:Opossums; D000644:Quaternary Ammonium Compounds / Q000032:analysis / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology
chemical_list: D005971:Glutamates; D006360:Heat-Shock Proteins; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D000643:Ammonium Chloride; D005973:Glutamine; D001224:Aspartic Acid; D018698:Glutamic Acid; D007770:L-Lactate Dehydrogenase