Modulation by dantrolene of endotoxin-induced interleukin-10, tumour necrosis factor-α and nitric oxide production in vivo and in vitro

György Haskó, Csaba Szabo, Zoltán H. Németh, Balázs Lendvai, E. Sylvester Vizi

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

1. Intracellular calcium has been suggested to be an important mediator of the cellular response in endotoxaemia and shock. Dantrolene is an agent that interferes with intracellular calcium fluxes resulting in a decreased availability of calcium in the cytoplasm. Here we have investigated the effect of dantrolene on lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10), tumour necrosis factor-α (TNF-α), and nitric oxide (NO) in mice and in cultured RAW 264.7 macrophages in vitro. 2. In BALB/c mice, LPS-induced plasma IL-10 levels were significantly enhanced by pretreatment with dantrolene (20 mg kg-1, i.p.) (P < 0.005 at the 90 min time-point). On the other hand, dantrolene pretreatment suppressed circulating TNF-α and nitrite/nitrate (breakdown products of NO) concentrations. However, dantrolene had no effect on LPS-induced plasma interleukin-6 (IL-6) levels (67.22 ± 5.51 ng ml-1 in vehicle-pretreated mice and 62.22 ± 3.66 ng ml-1 in dantrolene-pretreated mice, n = 9). 3. Dantrolene inhibited TNF-α and NO production in C57BL/6 IL-10(+/+) mice, as well as in their IL-10 deficient counterparts (C57BL/6 IL-10(0/0)). 4. In RAW 264.7 macrophages, dantrolene (10-300 μM) reduced IL-10, TNF-α, and nitrite (breakdown product of NO) production elicited by LPS (10 μg ml-1). Dantrolene (300 μM) did not affect the LPS-induced nuclear translocation of transcription factor nuclear factor κB in these cells. 5. Although LPS failed to alter the intracellullar concentration of calcium in single macrophages loaded with Fura-2, dantrolene caused a significant decrease of the basal calcium level as determined 30 min after dantrolene treatment (P < 0.005). ATP (1 mM) caused a rapid rise in intracellular calcium levels in both dantrolene-pretreated and vehicle-pretreated cells. 6. These results indicate that unlike the secretion of TNF-α and NO, IL-10 production is differentially regulated in vitro and in vivo. The decrease of plasma levels of the pro-inflammatory mediators TNF-α and NO, and increase in circulating IL-10 concentrations by dantrolene suggest that this drug might offer a new therapeutic approach in inflammatory diseases and septic shock.

Original languageEnglish (US)
Pages (from-to)1099-1106
Number of pages8
JournalBritish Journal of Pharmacology
Volume124
Issue number6
DOIs
StatePublished - 1998
Externally publishedYes

Fingerprint

Dantrolene
Endotoxins
Interleukin-10
Nitric Oxide
Tumor Necrosis Factor-alpha
Lipopolysaccharides
Calcium
Macrophages
Nitrites
In Vitro Techniques
Endotoxemia
Fura-2
Septic Shock
Nitrates

Keywords

  • Interleukin-10
  • Intracellular calcium
  • Lipopolysaccharide
  • Nitric oxide
  • Tumour necrosis factor-α

ASJC Scopus subject areas

  • Pharmacology

Cite this

Modulation by dantrolene of endotoxin-induced interleukin-10, tumour necrosis factor-α and nitric oxide production in vivo and in vitro. / Haskó, György; Szabo, Csaba; Németh, Zoltán H.; Lendvai, Balázs; Vizi, E. Sylvester.

In: British Journal of Pharmacology, Vol. 124, No. 6, 1998, p. 1099-1106.

Research output: Contribution to journalArticle

Haskó, György ; Szabo, Csaba ; Németh, Zoltán H. ; Lendvai, Balázs ; Vizi, E. Sylvester. / Modulation by dantrolene of endotoxin-induced interleukin-10, tumour necrosis factor-α and nitric oxide production in vivo and in vitro. In: British Journal of Pharmacology. 1998 ; Vol. 124, No. 6. pp. 1099-1106.
@article{e889dd61dfab45ff9ccf40c85a938d31,
title = "Modulation by dantrolene of endotoxin-induced interleukin-10, tumour necrosis factor-α and nitric oxide production in vivo and in vitro",
abstract = "1. Intracellular calcium has been suggested to be an important mediator of the cellular response in endotoxaemia and shock. Dantrolene is an agent that interferes with intracellular calcium fluxes resulting in a decreased availability of calcium in the cytoplasm. Here we have investigated the effect of dantrolene on lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10), tumour necrosis factor-α (TNF-α), and nitric oxide (NO) in mice and in cultured RAW 264.7 macrophages in vitro. 2. In BALB/c mice, LPS-induced plasma IL-10 levels were significantly enhanced by pretreatment with dantrolene (20 mg kg-1, i.p.) (P < 0.005 at the 90 min time-point). On the other hand, dantrolene pretreatment suppressed circulating TNF-α and nitrite/nitrate (breakdown products of NO) concentrations. However, dantrolene had no effect on LPS-induced plasma interleukin-6 (IL-6) levels (67.22 ± 5.51 ng ml-1 in vehicle-pretreated mice and 62.22 ± 3.66 ng ml-1 in dantrolene-pretreated mice, n = 9). 3. Dantrolene inhibited TNF-α and NO production in C57BL/6 IL-10(+/+) mice, as well as in their IL-10 deficient counterparts (C57BL/6 IL-10(0/0)). 4. In RAW 264.7 macrophages, dantrolene (10-300 μM) reduced IL-10, TNF-α, and nitrite (breakdown product of NO) production elicited by LPS (10 μg ml-1). Dantrolene (300 μM) did not affect the LPS-induced nuclear translocation of transcription factor nuclear factor κB in these cells. 5. Although LPS failed to alter the intracellullar concentration of calcium in single macrophages loaded with Fura-2, dantrolene caused a significant decrease of the basal calcium level as determined 30 min after dantrolene treatment (P < 0.005). ATP (1 mM) caused a rapid rise in intracellular calcium levels in both dantrolene-pretreated and vehicle-pretreated cells. 6. These results indicate that unlike the secretion of TNF-α and NO, IL-10 production is differentially regulated in vitro and in vivo. The decrease of plasma levels of the pro-inflammatory mediators TNF-α and NO, and increase in circulating IL-10 concentrations by dantrolene suggest that this drug might offer a new therapeutic approach in inflammatory diseases and septic shock.",
keywords = "Interleukin-10, Intracellular calcium, Lipopolysaccharide, Nitric oxide, Tumour necrosis factor-α",
author = "Gy{\"o}rgy Hask{\'o} and Csaba Szabo and N{\'e}meth, {Zolt{\'a}n H.} and Bal{\'a}zs Lendvai and Vizi, {E. Sylvester}",
year = "1998",
doi = "10.1038/sj.bjp.0701934",
language = "English (US)",
volume = "124",
pages = "1099--1106",
journal = "British Journal of Pharmacology",
issn = "0007-1188",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Modulation by dantrolene of endotoxin-induced interleukin-10, tumour necrosis factor-α and nitric oxide production in vivo and in vitro

AU - Haskó, György

AU - Szabo, Csaba

AU - Németh, Zoltán H.

AU - Lendvai, Balázs

AU - Vizi, E. Sylvester

PY - 1998

Y1 - 1998

N2 - 1. Intracellular calcium has been suggested to be an important mediator of the cellular response in endotoxaemia and shock. Dantrolene is an agent that interferes with intracellular calcium fluxes resulting in a decreased availability of calcium in the cytoplasm. Here we have investigated the effect of dantrolene on lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10), tumour necrosis factor-α (TNF-α), and nitric oxide (NO) in mice and in cultured RAW 264.7 macrophages in vitro. 2. In BALB/c mice, LPS-induced plasma IL-10 levels were significantly enhanced by pretreatment with dantrolene (20 mg kg-1, i.p.) (P < 0.005 at the 90 min time-point). On the other hand, dantrolene pretreatment suppressed circulating TNF-α and nitrite/nitrate (breakdown products of NO) concentrations. However, dantrolene had no effect on LPS-induced plasma interleukin-6 (IL-6) levels (67.22 ± 5.51 ng ml-1 in vehicle-pretreated mice and 62.22 ± 3.66 ng ml-1 in dantrolene-pretreated mice, n = 9). 3. Dantrolene inhibited TNF-α and NO production in C57BL/6 IL-10(+/+) mice, as well as in their IL-10 deficient counterparts (C57BL/6 IL-10(0/0)). 4. In RAW 264.7 macrophages, dantrolene (10-300 μM) reduced IL-10, TNF-α, and nitrite (breakdown product of NO) production elicited by LPS (10 μg ml-1). Dantrolene (300 μM) did not affect the LPS-induced nuclear translocation of transcription factor nuclear factor κB in these cells. 5. Although LPS failed to alter the intracellullar concentration of calcium in single macrophages loaded with Fura-2, dantrolene caused a significant decrease of the basal calcium level as determined 30 min after dantrolene treatment (P < 0.005). ATP (1 mM) caused a rapid rise in intracellular calcium levels in both dantrolene-pretreated and vehicle-pretreated cells. 6. These results indicate that unlike the secretion of TNF-α and NO, IL-10 production is differentially regulated in vitro and in vivo. The decrease of plasma levels of the pro-inflammatory mediators TNF-α and NO, and increase in circulating IL-10 concentrations by dantrolene suggest that this drug might offer a new therapeutic approach in inflammatory diseases and septic shock.

AB - 1. Intracellular calcium has been suggested to be an important mediator of the cellular response in endotoxaemia and shock. Dantrolene is an agent that interferes with intracellular calcium fluxes resulting in a decreased availability of calcium in the cytoplasm. Here we have investigated the effect of dantrolene on lipopolysaccharide (LPS)-induced production of interleukin-10 (IL-10), tumour necrosis factor-α (TNF-α), and nitric oxide (NO) in mice and in cultured RAW 264.7 macrophages in vitro. 2. In BALB/c mice, LPS-induced plasma IL-10 levels were significantly enhanced by pretreatment with dantrolene (20 mg kg-1, i.p.) (P < 0.005 at the 90 min time-point). On the other hand, dantrolene pretreatment suppressed circulating TNF-α and nitrite/nitrate (breakdown products of NO) concentrations. However, dantrolene had no effect on LPS-induced plasma interleukin-6 (IL-6) levels (67.22 ± 5.51 ng ml-1 in vehicle-pretreated mice and 62.22 ± 3.66 ng ml-1 in dantrolene-pretreated mice, n = 9). 3. Dantrolene inhibited TNF-α and NO production in C57BL/6 IL-10(+/+) mice, as well as in their IL-10 deficient counterparts (C57BL/6 IL-10(0/0)). 4. In RAW 264.7 macrophages, dantrolene (10-300 μM) reduced IL-10, TNF-α, and nitrite (breakdown product of NO) production elicited by LPS (10 μg ml-1). Dantrolene (300 μM) did not affect the LPS-induced nuclear translocation of transcription factor nuclear factor κB in these cells. 5. Although LPS failed to alter the intracellullar concentration of calcium in single macrophages loaded with Fura-2, dantrolene caused a significant decrease of the basal calcium level as determined 30 min after dantrolene treatment (P < 0.005). ATP (1 mM) caused a rapid rise in intracellular calcium levels in both dantrolene-pretreated and vehicle-pretreated cells. 6. These results indicate that unlike the secretion of TNF-α and NO, IL-10 production is differentially regulated in vitro and in vivo. The decrease of plasma levels of the pro-inflammatory mediators TNF-α and NO, and increase in circulating IL-10 concentrations by dantrolene suggest that this drug might offer a new therapeutic approach in inflammatory diseases and septic shock.

KW - Interleukin-10

KW - Intracellular calcium

KW - Lipopolysaccharide

KW - Nitric oxide

KW - Tumour necrosis factor-α

UR - http://www.scopus.com/inward/record.url?scp=0031853543&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031853543&partnerID=8YFLogxK

U2 - 10.1038/sj.bjp.0701934

DO - 10.1038/sj.bjp.0701934

M3 - Article

VL - 124

SP - 1099

EP - 1106

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 6

ER -