title: An in vitro model to consider the effect of 2 mM glutamine and KNK437 on endotoxin-stimulated release of heat shock protein 70 and inflammatory mediators.
abstract: OBJECTIVE
  Glutamine has been shown to promote the release of heat shock protein 70 (HSP70) both within experimental in vitro models of sepsis and in adults with septic shock. This study aimed to investigate the effects of 2 mM glutamine and an inhibitor of HSP70 (KNK437) on the release of HSP70 and inflammatory mediators in healthy adult volunteers.
  
  
  METHODS
  An in vitro whole blood endotoxin stimulation assay was used.
  
  
  RESULTS
  The addition of 2 mM glutamine significantly increased HSP70 levels over time (P < 0.05). HSP70 release had a positive correlation at 4 h with IL-1 β (r = 0.51, P = 0.03) and an inverse correlation with TNF-α (r = -0.56, P = 0.02) and IL-8 levels (r = -0.52, P = 0.03), and there were no significant correlations between HSP70 and IL6 or IL-10 or glutamine. Glutamine supplementation significantly (P < 0.05) attenuated the release of IL-10 at 4 h and IL-8 at 24 h, compared with conditions without glutamine. In endotoxin-stimulated blood there were no significant differences in the release of IL-6, TNF-α, and IL-1 β with glutamine supplementation at 4 and 24 h. However, glutamine supplementation (2 mM) appeared to attenuate the release of inflammatory mediators (IL-1 β, IL-6, TNF-α), although this effect was not statistically significant. The addition of KNK437, a HSP70 inhibitor, significantly diminished HSP70 release, which resulted in lower levels of inflammatory mediators (P < 0.05).
  
  
  CONCLUSIONS
  Glutamine supplementation promotes HSP70 release in an experimental model of sepsis. After the addition of KNK437, the effects of glutamine on HSP70 and inflammatory mediator release appear to be lost, suggesting that HSP70 in part orchestrates the inflammatory mediator response to sepsis. The clinical implications require further investigation.
authors.affiliation: Department of Paediatrics, Imperial College London, London, United Kingdom. Electronic address: luise.marino@uhs.nhs.uk.
authors.forename: Luise V
authors.initials: LV
authors.lastname: Marino
authors.affiliation: Department of Paediatrics, School of Clinical Medicine, Cambridge University, Cambridge, United Kingdom.
authors.forename: Nazima
authors.initials: N
authors.lastname: Pathan
authors.affiliation: Department of Gastroenterology, Great Ormond Street Hospital for Sick Children, London, United Kingdom.
authors.forename: Rosan W
authors.initials: RW
authors.lastname: Meyer
authors.affiliation: Department of Paediatrics, Imperial College London, London, United Kingdom.
authors.forename: Victoria J
authors.initials: VJ
authors.lastname: Wright
authors.affiliation: Department of Paediatrics, Imperial College London, London, United Kingdom.
authors.forename: Parviz
authors.initials: P
authors.lastname: Habibi
mesh_terms: D000328:Adult; D001559:Benzhydryl Compounds / Q000494:pharmacology*; D015992:Body Mass Index; D019587:Dietary Supplements; D004731:Endotoxins / Q000633:toxicity*; D005973:Glutamine / Q000494:pharmacology*; D018840:HSP70 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000097:blood*; D006801:Humans; D016753:Interleukin-10 / Q000097:blood; D053583:Interleukin-1beta / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D016209:Interleukin-8 / Q000097:blood; D008875:Middle Aged; D011760:Pyrrolidinones / Q000494:pharmacology*; D012772:Shock, Septic / Q000097:blood / Q000188:drug therapy; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D001559:Benzhydryl Compounds; C508606:CXCL8 protein, human; D004731:Endotoxins; D018840:HSP70 Heat-Shock Proteins; C508609:IL10 protein, human; C508585:IL1B protein, human; C508600:IL6 protein, human; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; C410027:KNK 437; D011760:Pyrrolidinones; D014409:Tumor Necrosis Factor-alpha; D005973:Glutamine; D016753:Interleukin-10
keywords: Glutamine; HSP70; Inflammatory mediators; KNK437; Sepsis