The Effect of Hypertonic Saline on mRNA of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Polymorphonuclear Cells

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Abstract

Background: Hypertonic saline is often used to resuscitate patients experiencing shock. In such conditions, polymorphonuclear cells and Toll-like receptors (TLRs) form an essential part of early induced innate immunity. Objective: To investigate the immunomodulatory effect of hypertonic saline on polymorphonuclear cells by evaluating the changes in TLR-4 receptors and proinflammatory cytokines. Methods: Polymorphonuclear cells were isolated from whole blood using Polymorphprep (Axis-Shield, Oslo, Norway). The isolated polymorphonuclear cells were plated at a density of 1 × 106cells/mL in 6-well flat-bottomed culture plates and were stimulated with 1 μg/mL lipopolysaccharide or N-formyl-methionyl-leucyl-phenylalanine. The stimulated polymorphonuclear cells were cultured in hypertonic saline at 10, 20, or 40 mmol/L above isotonicity. After that, the changes in TLR-4 and cytokines were measured by quantitative real-time polymerase chain reaction and flow cytometry. Results: The level of TLR-4 mRNA expression decreased after stimulation with N-formyl-methionyl-leucyl-phenylalanine, but hypertonic saline did not affect the TLR-4 mRNA expression. TLR-4 mRNA expression was clearly induced upon stimulation with lipopolysaccharide, and the addition of hypertonic saline restored TLR-4 mRNA expression in polymorphonuclear cells. The interleukin-1β mRNA expression was decreased in the hypertonic environment. On the other hand, the tumor necrosis factor-α value was not influenced by the addition of hypertonic saline. Conclusions: Hypertonic saline has an immunomodulatory effect on polymorphonuclear cells through the TLR-4 pathway, and the interleukin-1β-associated pathway is influenced more by hypertonic saline than is the tumor necrosis factor-α-associated pathway.

Original languageEnglish
Pages (from-to)58-62
Number of pages5
JournalCurrent Therapeutic Research - Clinical and Experimental
Volume76
DOIs
Publication statusPublished - 2014 Dec 1

Fingerprint

Toll-Like Receptor 4
Lipopolysaccharides
Cytokines
Messenger RNA
N-Formylmethionine Leucyl-Phenylalanine
Interleukin-1
Tumor Necrosis Factor-alpha
Cytokine Receptors
Toll-Like Receptors
Norway
Innate Immunity
Real-Time Polymerase Chain Reaction
Cultured Cells
Shock
Flow Cytometry

Keywords

  • Cytokine
  • Hypertonic saline solution
  • Immune
  • Inflammation
  • Neutrophil
  • Toll-like receptor 4

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)

Cite this

@article{583d077e3e4546c384e4047b5c17f574,
title = "The Effect of Hypertonic Saline on mRNA of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Polymorphonuclear Cells",
abstract = "Background: Hypertonic saline is often used to resuscitate patients experiencing shock. In such conditions, polymorphonuclear cells and Toll-like receptors (TLRs) form an essential part of early induced innate immunity. Objective: To investigate the immunomodulatory effect of hypertonic saline on polymorphonuclear cells by evaluating the changes in TLR-4 receptors and proinflammatory cytokines. Methods: Polymorphonuclear cells were isolated from whole blood using Polymorphprep (Axis-Shield, Oslo, Norway). The isolated polymorphonuclear cells were plated at a density of 1 × 106cells/mL in 6-well flat-bottomed culture plates and were stimulated with 1 μg/mL lipopolysaccharide or N-formyl-methionyl-leucyl-phenylalanine. The stimulated polymorphonuclear cells were cultured in hypertonic saline at 10, 20, or 40 mmol/L above isotonicity. After that, the changes in TLR-4 and cytokines were measured by quantitative real-time polymerase chain reaction and flow cytometry. Results: The level of TLR-4 mRNA expression decreased after stimulation with N-formyl-methionyl-leucyl-phenylalanine, but hypertonic saline did not affect the TLR-4 mRNA expression. TLR-4 mRNA expression was clearly induced upon stimulation with lipopolysaccharide, and the addition of hypertonic saline restored TLR-4 mRNA expression in polymorphonuclear cells. The interleukin-1β mRNA expression was decreased in the hypertonic environment. On the other hand, the tumor necrosis factor-α value was not influenced by the addition of hypertonic saline. Conclusions: Hypertonic saline has an immunomodulatory effect on polymorphonuclear cells through the TLR-4 pathway, and the interleukin-1β-associated pathway is influenced more by hypertonic saline than is the tumor necrosis factor-α-associated pathway.",
keywords = "Cytokine, Hypertonic saline solution, Immune, Inflammation, Neutrophil, Toll-like receptor 4",
author = "Choi, {Sung Hyuk} and Young-Hoon Yoon and Jung-Youn Kim and Sungwoo Moon and Cho, {Young Duck} and Yeom, {Ji Won}",
year = "2014",
month = "12",
day = "1",
doi = "10.1016/j.curtheres.2014.06.003",
language = "English",
volume = "76",
pages = "58--62",
journal = "Current Therapeutic Research - Clinical and Experimental",
issn = "0011-393X",
publisher = "Excerpta Medica",

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TY - JOUR

T1 - The Effect of Hypertonic Saline on mRNA of Proinflammatory Cytokines in Lipopolysaccharide-Stimulated Polymorphonuclear Cells

AU - Choi, Sung Hyuk

AU - Yoon, Young-Hoon

AU - Kim, Jung-Youn

AU - Moon, Sungwoo

AU - Cho, Young Duck

AU - Yeom, Ji Won

PY - 2014/12/1

Y1 - 2014/12/1

N2 - Background: Hypertonic saline is often used to resuscitate patients experiencing shock. In such conditions, polymorphonuclear cells and Toll-like receptors (TLRs) form an essential part of early induced innate immunity. Objective: To investigate the immunomodulatory effect of hypertonic saline on polymorphonuclear cells by evaluating the changes in TLR-4 receptors and proinflammatory cytokines. Methods: Polymorphonuclear cells were isolated from whole blood using Polymorphprep (Axis-Shield, Oslo, Norway). The isolated polymorphonuclear cells were plated at a density of 1 × 106cells/mL in 6-well flat-bottomed culture plates and were stimulated with 1 μg/mL lipopolysaccharide or N-formyl-methionyl-leucyl-phenylalanine. The stimulated polymorphonuclear cells were cultured in hypertonic saline at 10, 20, or 40 mmol/L above isotonicity. After that, the changes in TLR-4 and cytokines were measured by quantitative real-time polymerase chain reaction and flow cytometry. Results: The level of TLR-4 mRNA expression decreased after stimulation with N-formyl-methionyl-leucyl-phenylalanine, but hypertonic saline did not affect the TLR-4 mRNA expression. TLR-4 mRNA expression was clearly induced upon stimulation with lipopolysaccharide, and the addition of hypertonic saline restored TLR-4 mRNA expression in polymorphonuclear cells. The interleukin-1β mRNA expression was decreased in the hypertonic environment. On the other hand, the tumor necrosis factor-α value was not influenced by the addition of hypertonic saline. Conclusions: Hypertonic saline has an immunomodulatory effect on polymorphonuclear cells through the TLR-4 pathway, and the interleukin-1β-associated pathway is influenced more by hypertonic saline than is the tumor necrosis factor-α-associated pathway.

AB - Background: Hypertonic saline is often used to resuscitate patients experiencing shock. In such conditions, polymorphonuclear cells and Toll-like receptors (TLRs) form an essential part of early induced innate immunity. Objective: To investigate the immunomodulatory effect of hypertonic saline on polymorphonuclear cells by evaluating the changes in TLR-4 receptors and proinflammatory cytokines. Methods: Polymorphonuclear cells were isolated from whole blood using Polymorphprep (Axis-Shield, Oslo, Norway). The isolated polymorphonuclear cells were plated at a density of 1 × 106cells/mL in 6-well flat-bottomed culture plates and were stimulated with 1 μg/mL lipopolysaccharide or N-formyl-methionyl-leucyl-phenylalanine. The stimulated polymorphonuclear cells were cultured in hypertonic saline at 10, 20, or 40 mmol/L above isotonicity. After that, the changes in TLR-4 and cytokines were measured by quantitative real-time polymerase chain reaction and flow cytometry. Results: The level of TLR-4 mRNA expression decreased after stimulation with N-formyl-methionyl-leucyl-phenylalanine, but hypertonic saline did not affect the TLR-4 mRNA expression. TLR-4 mRNA expression was clearly induced upon stimulation with lipopolysaccharide, and the addition of hypertonic saline restored TLR-4 mRNA expression in polymorphonuclear cells. The interleukin-1β mRNA expression was decreased in the hypertonic environment. On the other hand, the tumor necrosis factor-α value was not influenced by the addition of hypertonic saline. Conclusions: Hypertonic saline has an immunomodulatory effect on polymorphonuclear cells through the TLR-4 pathway, and the interleukin-1β-associated pathway is influenced more by hypertonic saline than is the tumor necrosis factor-α-associated pathway.

KW - Cytokine

KW - Hypertonic saline solution

KW - Immune

KW - Inflammation

KW - Neutrophil

KW - Toll-like receptor 4

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U2 - 10.1016/j.curtheres.2014.06.003

DO - 10.1016/j.curtheres.2014.06.003

M3 - Article

VL - 76

SP - 58

EP - 62

JO - Current Therapeutic Research - Clinical and Experimental

JF - Current Therapeutic Research - Clinical and Experimental

SN - 0011-393X

ER -