Mechanism of inhibitory action of ethanol on inducible nitric oxide synthesis in macrophages

Wakabayashi, Ichiro; Negoro, Munetaka

doi:10.1007/s00210-002-0647-6

Cited by 3 publications

(4 citation statements)

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“…Investigating the effects of ethanol on HeLa cells it was found that concentrations of 5 % and above compromised the cell viability (Forman et al 1999). The influence of ethanol on cell viability of RAW 264.7 cells was evaluated by the trypan blue staining method and it was found that the cell viability was not affected at concentrations up to 600 mM (*2.8 %) (Wakabayashi and Negoro 2002). In the present study RAW 264.7 cells were found to tolerate concentrations of ethanol of 1 % while the response was greatly attenuated at 5 %.…”

Section: Discussionmentioning

confidence: 99%

Considerations regarding use of solvents in in vitro cell based assays

et al. 2013

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Cell culture systems are widely used for the investigation of in vitro immunomodulatory effects of medicines and natural products. Since many pharmacological relevant compounds are water-insoluble, solvents are frequently used in cell based assays. Although many reports describe the cellular effects of solvents at high concentrations, only a few relate the effects of solvents used at low concentrations. In this report we investigate the interference of three commonly used solvents: Dimethyl sulfoxide (DMSO), ethanol and β-cyclodextrin with five different cell culture systems. The effects of the solvents are investigated in relation to the cellular production of interleukin (IL)-6 or reactive oxygen species (ROS) after lipopolysaccharide (LPS) stimulation. We show that DMSO above 1 % reduces readout parameters in all cell types but more interestingly the 0.25 and 0.5 % solutions induce inhibitory effects in some cell types and stimulatory effects in others. We also found that LPS induced ROS production was more affected than the IL-6 production in the presence of ethanol. Finally we showed that β-cyclodextrin at the investigated concentrations did not have any effect on the LPS induced IL-6 production and only minor effects on the ROS production. We conclude that the effects induced by solvents even at low concentrations are highly relevant for the interpretation of immunomodulatory effects evaluated in cell assays. Furthermore, these results show the importance of keeping solvent concentrations constant in serial dilution of any compound investigated in cell based assays.

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Section: Discussionmentioning

confidence: 99%

Considerations regarding use of solvents in in vitro cell based assays

et al. 2013

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“…2005) diminished iNOS synthesis (Zhang et al. 1998), possibly because of the inhibition of STAT1 activation (Wakabayashi and Negoro 2002) or of the phospholipase d ‐tyrosine kinase pathway involved in the phosphorylation and activation of NADPH oxidase (Greenberg et al. 1998).…”

Section: Discussionmentioning

confidence: 99%

Neuro‐inflammation induced in the hippocampus of ‘binge drinking’ rats may be mediated by elevated extracellular glutamate content

Ward

Colivicchi

Allen

et al. 2009

Journal of Neurochemistry

116

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The neuropathological and immune changes induced in the brain by ‘binge drinking’ have been investigated in a rat model. Evidence of neuro‐inflammation was identified in the ‘binge drinking’ rat model of alcohol abuse after 3 weeks of administration of 2 or 3 g/kg ethanol (EtOH), three times per day for two consecutive days, followed by 5 days of abstinence: Firstly, alveolar macrophages, isolated from these animals, showed significant increases in inducible nitric oxide synthase, as assayed by nitrite release, both before and after lipopolysaccaharide stimulation. Secondly, significant numbers of activated microglia were present in the dentate gyrus region of the hippocampus of the ‘binge drinking’ model, after major histocompatibility complex class II staining, by comparison with the control. Microdialysis studies in the ventral hippocampus identified a significant increase in the basal extracellular concentration of glutamate, in both the 2 and 3 g/kg administered ‘binge drinking’ rats. In contrast, no changes in the hippocampal extracellular concentrations, of GABA and taurine, or the dopamine and serotonin metabolites were observed under basal conditions. A further dose of EtOH induced a significant decrease in the concentrations of both 3,4‐dihydroxyphenylacetic acid and 5‐hydroxyindoleacetic acid, whereas glutamate, taurine and GABA levels were unaffected. There was no evidence that EtOH preference was initiated by the ‘binge drinking’ regimen. Our results suggest that the possible toxicity associated with ‘binge drinking’ maybe directed by the elevated glutamate levels, which in turn, activate phagocytic cells to release their inflammatory cytokines and chemokines, ultimately leading to neuro‐inflammation.

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“…In inflammatory contexts, there is increase in the secretion of soluble mediators such as pro‐inflammatory cytokines, including TNF‐α, IL‐1, IL‐6 and IL‐8. Activation of macrophage by pro‐inflammatory cytokines is an efficient pathway to activate iNOS leading to higher synthesis of nitric oxide (NO) directly involved in macrophage bactericidal activity and efficient phagocytosis [1, 3].…”

Section: Introductionmentioning

confidence: 99%

“…Indeed, most reports have focussed on early alterations induced by EtOH consumption or those observed following direct in vitro addition of EtOH over macrophages and lymphocyte cultures [4–7]. The immunological alterations already reported following EtOH administration describe mainly the suppression of inflammatory‐type cytokines, including reduced levels IL‐2 and IFN‐γ [8, 9], impairment of TNF‐α, IL‐1 and IL‐6 production by human macrophages and murine Kupffer cells [10–12], and inhibition of iNOS induction at the transcriptional level in macrophage [3].…”

Section: Introductionmentioning

confidence: 99%

The Long‐term Impaired Macrophages Functions are Already Observed Early after High‐dose Ethanol Administration

et al. 2008

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Herein, we described an experimental model of high‐dose ethanol (EtOH) administration, able to induce in vitro impairment in macrophage phagocytic capacity, already observed at 24 h after the last EtOH administration. This phenomenon was characterized by enlarged time required for adhesion and internalization events. Parallel studies documented an overall impaired production of interleukin (IL)‐6 and nitric oxide (NO) production by peritoneal macrophages in EtOH‐treated mice following interferon (IFN)‐γ and lipopolysaccharide (LPS) stimuli. Although the impaired IL‐6 response could not be restored by any of the experimental conditions tested, the lower NO response to INF‐γ and LPS was overturned by simultaneous IFN‐γ/LPS stimuli. It was interesting to notice that high‐dose EtOH administration drives peritoneal macrophages towards long‐term impairment in phagocytosis capacity with slower adhesion time, but with no impact on the time required for internalization. Moreover, 30 days after the last EtOH administration, lower IL‐6 response to INF‐γ and impaired NO production were still observed in response to IFN‐γ/LPS stimuli, with the IL‐6 response to IFN‐γ being restored by IFN‐γ/LPS stimuli. Histological studies showed that high‐dose EtOH administration led to long‐term in vivo impairment of antigen‐clearance following OVA‐driven delayed‐type‐hypersensitivity induction, characterized by the presence of a large amount of unprocessed OVA surrounded by dermal inflammatory infiltrate, suggesting defective activity of antigen‐presenting cells. Together, these findings supported our hypothesis that the poor antigen clearance in vivo may be related to the impaired macrophage function in vitro. These observations in the murine experimental model may reflect some of the consequences of EtOH consumption by humans.

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Mechanism of inhibitory action of ethanol on inducible nitric oxide synthesis in macrophages

Cited by 3 publications

References 0 publications

Considerations regarding use of solvents in in vitro cell based assays

Considerations regarding use of solvents in in vitro cell based assays

Neuro‐inflammation induced in the hippocampus of ‘binge drinking’ rats may be mediated by elevated extracellular glutamate content

The Long‐term Impaired Macrophages Functions are Already Observed Early after High‐dose Ethanol Administration

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