cDNA profiling in leukocytes exposed to hypertonic resuscitation fluids

Vadim Gushchin, Hasan B. Alam, Peter M Rhee, John R. Kirkpatrick, Elena Koustova

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background Resuscitative fluids induce distinctive changes in leukocyte functions: incubation with colloid Dextran increases production of reactive oxygen species and adhesion, whereas exposure to hypertonic saline (HTS) inhibits "oxidative burst" and phagocytosis. In hypertonic saline Dextran (HTD), the hypertonic component determines the leukocyte functional behavior and subsequently activation response. We investigated whether leukocyte gene expression is analogously affected. Methods Whole blood from eight volunteers was diluted and incubated for 30 min at 37oC in 6. 0% Dextran-70, 7.5% HTS, and 7.5% HTD. Total leukocyte RNA was extracted and used to synthesize biotinylated cDNA probes. Each probe was individually hybridized to a cDNA array to simultaneously measure the expression of 23 genes involved in inflammation, cell migration, and apoptosis. Results Leukocytes incubated with Dextran-70 demonstrated greater than a 6-fold (p < 0.05) increase in the expression of interleukin-8, growth-regulated oncogenes α and β, L-selectin, superoxide dismutase, tumor necrosis factor-α (TNF-α), and mitogen-activated protein kinase 3. The expression profile induced by HTS was not significantly different from that of unstimulated blood, except for prominent induction of only three genes. HTD attenuated the expression of Dextran-70 upregulated genes, although the level of their expression was higher than in HTS-treated leukocytes. Conclusions Hypertonic resuscitation fluids diminish the expression of immune activation-associated genes. Hypertonic component of HTD determines the leukocyte gene expression profile.

Original languageEnglish (US)
Pages (from-to)426-432
Number of pages7
JournalJournal of the American College of Surgeons
Volume197
Issue number3
DOIs
StatePublished - Sep 2003
Externally publishedYes

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dextran - saline drug combination
Resuscitation
Leukocytes
Complementary DNA
Dextrans
Gene Expression
L-Selectin
Mitogen-Activated Protein Kinase 3
Respiratory Burst
Colloids
Oligonucleotide Array Sequence Analysis
Interleukin-8
Oncogenes
Transcriptome
Phagocytosis
Transcriptional Activation
Genes
Superoxide Dismutase
Cell Movement
Volunteers

Keywords

  • DEX
  • Dextran-70
  • GAPDH
  • Glyceraldehyde-3-phosphate dehydrogenase
  • GRO
  • Growth-regulated oncogenes
  • HTD
  • HTS
  • Hypertonic saline
  • Hypertonic saline Dextran
  • IL
  • Interleukin
  • PMN
  • Polymorphonuclear
  • TNF
  • Tumor necrosis factor
  • WB
  • Whole blood

ASJC Scopus subject areas

  • Surgery

Cite this

cDNA profiling in leukocytes exposed to hypertonic resuscitation fluids. / Gushchin, Vadim; Alam, Hasan B.; Rhee, Peter M; Kirkpatrick, John R.; Koustova, Elena.

In: Journal of the American College of Surgeons, Vol. 197, No. 3, 09.2003, p. 426-432.

Research output: Contribution to journalArticle

Gushchin, Vadim ; Alam, Hasan B. ; Rhee, Peter M ; Kirkpatrick, John R. ; Koustova, Elena. / cDNA profiling in leukocytes exposed to hypertonic resuscitation fluids. In: Journal of the American College of Surgeons. 2003 ; Vol. 197, No. 3. pp. 426-432.
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AU - Gushchin, Vadim

AU - Alam, Hasan B.

AU - Rhee, Peter M

AU - Kirkpatrick, John R.

AU - Koustova, Elena

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N2 - Background Resuscitative fluids induce distinctive changes in leukocyte functions: incubation with colloid Dextran increases production of reactive oxygen species and adhesion, whereas exposure to hypertonic saline (HTS) inhibits "oxidative burst" and phagocytosis. In hypertonic saline Dextran (HTD), the hypertonic component determines the leukocyte functional behavior and subsequently activation response. We investigated whether leukocyte gene expression is analogously affected. Methods Whole blood from eight volunteers was diluted and incubated for 30 min at 37oC in 6. 0% Dextran-70, 7.5% HTS, and 7.5% HTD. Total leukocyte RNA was extracted and used to synthesize biotinylated cDNA probes. Each probe was individually hybridized to a cDNA array to simultaneously measure the expression of 23 genes involved in inflammation, cell migration, and apoptosis. Results Leukocytes incubated with Dextran-70 demonstrated greater than a 6-fold (p < 0.05) increase in the expression of interleukin-8, growth-regulated oncogenes α and β, L-selectin, superoxide dismutase, tumor necrosis factor-α (TNF-α), and mitogen-activated protein kinase 3. The expression profile induced by HTS was not significantly different from that of unstimulated blood, except for prominent induction of only three genes. HTD attenuated the expression of Dextran-70 upregulated genes, although the level of their expression was higher than in HTS-treated leukocytes. Conclusions Hypertonic resuscitation fluids diminish the expression of immune activation-associated genes. Hypertonic component of HTD determines the leukocyte gene expression profile.

AB - Background Resuscitative fluids induce distinctive changes in leukocyte functions: incubation with colloid Dextran increases production of reactive oxygen species and adhesion, whereas exposure to hypertonic saline (HTS) inhibits "oxidative burst" and phagocytosis. In hypertonic saline Dextran (HTD), the hypertonic component determines the leukocyte functional behavior and subsequently activation response. We investigated whether leukocyte gene expression is analogously affected. Methods Whole blood from eight volunteers was diluted and incubated for 30 min at 37oC in 6. 0% Dextran-70, 7.5% HTS, and 7.5% HTD. Total leukocyte RNA was extracted and used to synthesize biotinylated cDNA probes. Each probe was individually hybridized to a cDNA array to simultaneously measure the expression of 23 genes involved in inflammation, cell migration, and apoptosis. Results Leukocytes incubated with Dextran-70 demonstrated greater than a 6-fold (p < 0.05) increase in the expression of interleukin-8, growth-regulated oncogenes α and β, L-selectin, superoxide dismutase, tumor necrosis factor-α (TNF-α), and mitogen-activated protein kinase 3. The expression profile induced by HTS was not significantly different from that of unstimulated blood, except for prominent induction of only three genes. HTD attenuated the expression of Dextran-70 upregulated genes, although the level of their expression was higher than in HTS-treated leukocytes. Conclusions Hypertonic resuscitation fluids diminish the expression of immune activation-associated genes. Hypertonic component of HTD determines the leukocyte gene expression profile.

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KW - Tumor necrosis factor

KW - WB

KW - Whole blood

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