Role of sodium in intracellular calcium elevation and leukotriene B 4 formation by receptor-mediated activation of human neutrophils

Alberto Tedeschi, Paola Ciceri, Simona Zarini, Maurizio Lorini, Manuela Di Donato, Simonetta Nicosia, Antonio Miadonna, Angelo Sala

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The role of Na+ and Na+ exchangers in intracellular Ca2+ elevation and leukotriene B4 (LTBs) formation was investigated in granulocyte macrophage colony-stimulating factor (GM-CSF)-primed, fMLP-stimulated human neutrophils. Isotonic substitution of extracellular Na+ with N-methyl-D-glucamine+ (NMDG +) resulted in over 85% inhibition of the LTBs generation observed (from 14.1±0.9pmol/106 neutrophils to 1.7±1.0pmol/ 106 neutrophils at 0.3μM fMLP). Isotonic substitution of Na + with NMDG+ also induced a significant inhibition of fMLP-induced rise in cytosolic Ca2+ concentration ([Ca 2+]i) (from 2.17- to 0.78-fold increase over basal levels). Pretreatment with an inhibitor of the Na+/Ca2+ exchanger (benzamil) did not inhibit either [Ca2+]i rise or LTBs production, indicating that the observed effects of extracellular Na+-deprivation were unrelated to the Na+/Ca2+ exchanger in receptor-mediated Ca2+ influx, as previously hypothesized. LTBs production by thapsigargin-activated neutrophils was not affected by Na+ depletion, but was totally abolished in the presence of EGTA, suggesting that store depletion-driven extracellular Ca2+ influx is required for leukotriene synthesis and that this process is independent of Na+-deprivation. Exposure to Na+-free medium for the time of GM-CSF priming led to a significant decrease of intracellular pH values, suggesting a role of the Na+/H+ exchanger in intracellular Na+ depletion. Reducing the time of Na+-deprivation totally reversed the observed effect on LTBs production, resulting in enhanced, rather than inhibited, formation of LTBs. These results indicate that LTBs generation and [Ca2+]i rise in human neutrophils primed by GM-CSF and stimulated with fMLP is dependent on intracellular Na+ concentration, and, at variance with previously published results, unrelated to the Ca2+ influx through the Na+/Ca2+ exchanger.

Original languageEnglish
Pages (from-to)385-393
Number of pages9
JournalBiochemical Pharmacology
Volume67
Issue number2
DOIs
Publication statusPublished - Jan 15 2004

Fingerprint

Neutrophil Activation
Leukotriene B4
Sodium
Chemical activation
Calcium
Neutrophils
Granulocyte-Macrophage Colony-Stimulating Factor
Substitution reactions
Sodium-Hydrogen Antiporter
Thapsigargin
Leukotrienes
Egtazic Acid

Keywords

  • Calcium ion
  • Leukotriene B
  • Neutrophils
  • Sodium ion

ASJC Scopus subject areas

  • Pharmacology

Cite this

Role of sodium in intracellular calcium elevation and leukotriene B 4 formation by receptor-mediated activation of human neutrophils. / Tedeschi, Alberto; Ciceri, Paola; Zarini, Simona; Lorini, Maurizio; Di Donato, Manuela; Nicosia, Simonetta; Miadonna, Antonio; Sala, Angelo.

In: Biochemical Pharmacology, Vol. 67, No. 2, 15.01.2004, p. 385-393.

Research output: Contribution to journalArticle

Tedeschi, A, Ciceri, P, Zarini, S, Lorini, M, Di Donato, M, Nicosia, S, Miadonna, A & Sala, A 2004, 'Role of sodium in intracellular calcium elevation and leukotriene B 4 formation by receptor-mediated activation of human neutrophils', Biochemical Pharmacology, vol. 67, no. 2, pp. 385-393. https://doi.org/10.1016/j.bcp.2003.09.019
Tedeschi, Alberto ; Ciceri, Paola ; Zarini, Simona ; Lorini, Maurizio ; Di Donato, Manuela ; Nicosia, Simonetta ; Miadonna, Antonio ; Sala, Angelo. / Role of sodium in intracellular calcium elevation and leukotriene B 4 formation by receptor-mediated activation of human neutrophils. In: Biochemical Pharmacology. 2004 ; Vol. 67, No. 2. pp. 385-393.
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AU - Lorini, Maurizio

AU - Di Donato, Manuela

AU - Nicosia, Simonetta

AU - Miadonna, Antonio

AU - Sala, Angelo

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N2 - The role of Na+ and Na+ exchangers in intracellular Ca2+ elevation and leukotriene B4 (LTBs) formation was investigated in granulocyte macrophage colony-stimulating factor (GM-CSF)-primed, fMLP-stimulated human neutrophils. Isotonic substitution of extracellular Na+ with N-methyl-D-glucamine+ (NMDG +) resulted in over 85% inhibition of the LTBs generation observed (from 14.1±0.9pmol/106 neutrophils to 1.7±1.0pmol/ 106 neutrophils at 0.3μM fMLP). Isotonic substitution of Na + with NMDG+ also induced a significant inhibition of fMLP-induced rise in cytosolic Ca2+ concentration ([Ca 2+]i) (from 2.17- to 0.78-fold increase over basal levels). Pretreatment with an inhibitor of the Na+/Ca2+ exchanger (benzamil) did not inhibit either [Ca2+]i rise or LTBs production, indicating that the observed effects of extracellular Na+-deprivation were unrelated to the Na+/Ca2+ exchanger in receptor-mediated Ca2+ influx, as previously hypothesized. LTBs production by thapsigargin-activated neutrophils was not affected by Na+ depletion, but was totally abolished in the presence of EGTA, suggesting that store depletion-driven extracellular Ca2+ influx is required for leukotriene synthesis and that this process is independent of Na+-deprivation. Exposure to Na+-free medium for the time of GM-CSF priming led to a significant decrease of intracellular pH values, suggesting a role of the Na+/H+ exchanger in intracellular Na+ depletion. Reducing the time of Na+-deprivation totally reversed the observed effect on LTBs production, resulting in enhanced, rather than inhibited, formation of LTBs. These results indicate that LTBs generation and [Ca2+]i rise in human neutrophils primed by GM-CSF and stimulated with fMLP is dependent on intracellular Na+ concentration, and, at variance with previously published results, unrelated to the Ca2+ influx through the Na+/Ca2+ exchanger.

AB - The role of Na+ and Na+ exchangers in intracellular Ca2+ elevation and leukotriene B4 (LTBs) formation was investigated in granulocyte macrophage colony-stimulating factor (GM-CSF)-primed, fMLP-stimulated human neutrophils. Isotonic substitution of extracellular Na+ with N-methyl-D-glucamine+ (NMDG +) resulted in over 85% inhibition of the LTBs generation observed (from 14.1±0.9pmol/106 neutrophils to 1.7±1.0pmol/ 106 neutrophils at 0.3μM fMLP). Isotonic substitution of Na + with NMDG+ also induced a significant inhibition of fMLP-induced rise in cytosolic Ca2+ concentration ([Ca 2+]i) (from 2.17- to 0.78-fold increase over basal levels). Pretreatment with an inhibitor of the Na+/Ca2+ exchanger (benzamil) did not inhibit either [Ca2+]i rise or LTBs production, indicating that the observed effects of extracellular Na+-deprivation were unrelated to the Na+/Ca2+ exchanger in receptor-mediated Ca2+ influx, as previously hypothesized. LTBs production by thapsigargin-activated neutrophils was not affected by Na+ depletion, but was totally abolished in the presence of EGTA, suggesting that store depletion-driven extracellular Ca2+ influx is required for leukotriene synthesis and that this process is independent of Na+-deprivation. Exposure to Na+-free medium for the time of GM-CSF priming led to a significant decrease of intracellular pH values, suggesting a role of the Na+/H+ exchanger in intracellular Na+ depletion. Reducing the time of Na+-deprivation totally reversed the observed effect on LTBs production, resulting in enhanced, rather than inhibited, formation of LTBs. These results indicate that LTBs generation and [Ca2+]i rise in human neutrophils primed by GM-CSF and stimulated with fMLP is dependent on intracellular Na+ concentration, and, at variance with previously published results, unrelated to the Ca2+ influx through the Na+/Ca2+ exchanger.

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