Endotoxin inhibits heat shock protein 70 (HSP70) expression in peripheral blood mononuclear cells of patients with severe sepsis

Schroeder, Stefan; Bischoff, Jenny; Lehmann, Lutz; Hering, R.; Spiegel, T. von; Putensen, Christian; Hoeft, Andreas; Stüber, Frank

doi:10.1007/s001340050786

Cited by 32 publications

(16 citation statements)

References 24 publications

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“…We also demonstrated that SA induced the expression of newly synthesized HSP70. Since there are a number of reports on the protective role of HSP70 in LPS-induced tissue injury (9,17,22,28), it was suggested that the prevention of LPS-induced injury by SA might be related to SA-induced heat shock response.…”

Section: Discussionmentioning

confidence: 99%

Cytoskeletal Alterations in Lipopolysaccharide-Induced Bovine Vascular Endothelial Cell Injury and Its Prevention by Sodium Arsenite

Chakravortty

Koide

Kato

et al. 2000

Clin Diagn Lab Immunol

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Morphological changes, especially cytoskeletal alterations, in lipopolysaccharide (LPS)-induced vascular endothelial cell injury were studied by using LPS-susceptible bovine aortic endothelial cells (BAEC). BAEC in cultures with LPS showed cell rounding, shrinking, and intercellular gap formation. In those cells, LPS caused the disorganization of actin, tubulin, and vimentin. LPS also induced a reduction in the F-actin pool and an elevation in the G-actin pool. Cytoskeletal disorganization affected transendothelial permeability across the endothelial monolayer. Pretreatment of BAEC with sodium arsenite (SA) prevented alterations in LPS-induced BAEC injury. However, posttreatment with SA had no protective effect on them. SA upregulated the expression of heat shock protein in the presence of LPS. The role of SA in prevention of LPS-induced BAEC injury is discussed.Bacterial lipopolysaccharide (LPS), an outer membrane component of gram-negative bacteria, has been shown to directly induce systemic injury of vascular endothelial cells and cause systemic inflammatory response syndrome or multiorgan failure (3). In an in vitro culture system, LPS was reported to induce the injury of bovine aortic endothelial cells (BAEC) directly in the absence of nonendothelial-cell-derived host mediators (11,13,20,23). The LPS-induced BAEC injury is accompanied by altered cell morphology, intercellular gap formation, and increased transendothelial permeability (12,21,24). It was possible that cytoskeletal alterations in LPS-induced BAEC injury were closely linked to intercellular gap formation and endothelial barrier dysfunction (12). However, there were few reports on detailed alterations of cytoskeleton in morphological changes of LPS-induced BAEC injury (12). Furthermore, knowledge regarding factors preventing the alterations in LPS-induced BAEC injury is very limited (15,16,26).Sodium arsenite (SA) is known to be a standard inducer of the heat shock response in vitro and can lead to heat shock protein (HSP) expression in vascular endothelial cells (4, 5). Several reports suggest that SA prevented LPS-induced endothelial cell injury via enhanced heat shock response (6, 27, 32). Therefore, it was of particular interest to determine if and how SA affected morphological changes in LPS-induced BAEC injury. In the present study, we examined the detailed cytoskeletal alterations in LPS-induced vascular endothelial cell injury by using LPS-susceptible BAEC and, furthermore, observed the effect of SA on them. Here we discuss the role of SA in the prevention of LPS-induced BAEC injury. MATERIALS AND METHODSMaterials. LPS from Escherichia coli O55:B5 was obtained from Sigma Chemical Co., St. Louis, Mo. LPS was dissolved at a concentration of 1 mg/ml in distilled water and diluted in culture medium for experiments. SA (Wako Pure Chemicals, Osaka, Japan) was dissolved at a concentration of 1 mM and diluted to 100 M in culture medium for experiments.Cell culture. BAEC were obtained from the Health Science Resource Bank (Tokyo, Japan) and maint...

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Section: Discussionmentioning

confidence: 99%

Cytoskeletal Alterations in Lipopolysaccharide-Induced Bovine Vascular Endothelial Cell Injury and Its Prevention by Sodium Arsenite

Chakravortty

Koide

Kato

et al. 2000

Clin Diagn Lab Immunol

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“…These results suggest that HSP70 influences the cytokine milieu release, mediating an optimal inflammatory response dependent on the level of stress [21], an effect supported by others [50,51].…”

Section: Discussionmentioning

confidence: 69%

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

et al. 2016

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Please cite this article as: Marino LV, Pathan N, Meyer R, Wright VJ, Habibi P, An in vitro model to consider the effect of 2mM glutamine and KNK437 on endotoxin stimulated release of HSP70 and inflammatory mediators, Nutrition (2015), doi: 10.1016/j.nut.2015.09.007. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Objective: Glutamine has been shown to promote heat shock protein 70 (HSP70) release both within 2 experimental in vitro models of sepsis and in adults with septic shock. 3Methods: An in vitro whole blood endotoxin stimulation assay was used to investigate the effects of 4 2mM glutamine and an inhibitor of HSP70 (KNK437) on the release of HSP70 and inflammatory 5 mediators in healthy adult volunteers. 6Results: 2mM glutamine significantly increased HSP70 levels over time (p<0.05). HSP70 release had a 7 positive correlation at 4 hours with IL-1β (r=0.51, p=0.03), an inverse correlation with TNF-α (r=-8 0.56, p=0.02) and IL-8 levels (r=-0.52, p=0.03), and there were no significant correlations between 9 HSP70 and IL6 or IL-10 or glutamine. Glutamine supplementation significantly (p<0.05) attenuated 10 the release of IL-10 at 4 hours and IL-8 at 24 hours, compared to conditions without glutamine. In 11 endotoxin stimulated blood there were no significant differences in the release of IL-6, TNF-α and IL-12 1β with glutamine supplementation at 4 and 24 hours. However, glutamine supplementation (2mM) 13 appeared to attenuate the release of inflammatory mediators (IL-1β, IL-6, TNF-α), although this 14 effect was not statistically significant. The addition of KNK 437, an HSP70 inhibitor significantly 15 diminished HSP70 release which resulted in lower levels of inflammatory mediators (p<0.05). 16Conclusion: Glutamine supplementation promotes HSP70 release in an experimental model of 17 sepsis. Following the addition of KNK437 the effects of glutamine on HSP70 and inflammatory 18 mediator release appear to be lost, suggesting the HSP70 in part orchestrates the inflammatory 19 mediator response to sepsis. The clinical implications require further investigation.

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“…It is likely that this GLN deficiency leads to a condition in which the patient is unable to induce HSP expression appropriately. Preliminary studies in immune cells taken from critically ill patients support this conclusion [14,15]. This would place the critically ill individual in a compromised position in which appropriate tissue protection, immune regulation, and preservation of metabolic function could be significantly impaired.…”

Section: Resultsmentioning

confidence: 75%

“…In this study, adenoviral transfection of HSP-70 into pulmonary epithelium ameliorated the occurrence of ARDS following sepsis. Clinical data reveal that the expression of HSP-70 is impaired in lymphocytes of patients with severe sepsis [14,15].…”

Section: Introductionmentioning

confidence: 99%

Glutamine: the first clinically relevant pharmacological regulator of heat shock protein expression?

Wischmeyer

2006

Current Opinion in Clinical Nutrition &Amp; Metabolic Care

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Currently, extensive data support glutamine as a gene level regulator of heat shock protein expression. Glutamine depletion, following critical illness/injury, is likely to lead to a state in which organisms are unable to induce heat shock proteins appropriately. Further, pharmacologic supplementation of glutamine potentiates the heat shock protein response prior to and following a stress. Pharmacologic trials utilizing glutamine to enhance heat shock proteins in humans are indicated.

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Endotoxin inhibits heat shock protein 70 (HSP70) expression in peripheral blood mononuclear cells of patients with severe sepsis

Cited by 32 publications

References 24 publications

Cytoskeletal Alterations in Lipopolysaccharide-Induced Bovine Vascular Endothelial Cell Injury and Its Prevention by Sodium Arsenite

Cytoskeletal Alterations in Lipopolysaccharide-Induced Bovine Vascular Endothelial Cell Injury and Its Prevention by Sodium Arsenite

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

Glutamine: the first clinically relevant pharmacological regulator of heat shock protein expression?

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