Effect of chronic ethanol administration on the in vitro production of proinflammatory cytokines by rat Kupffer cells in the presence of apoptotic cells

Benita L McVicker, Dean J. Tuma, Kusum Kharbanda, Jacy L. Kubik, Carol A Casey

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Background: Chronic ethanol consumption can lead to a variety of pathological consequences by as yet undefined mechanisms. Recently, it has been noted that alcohol-associated liver disease is often accompanied by morphological liver changes that include the increased production of apoptotic cells. Additionally, it has been demonstrated that hepatocellular uptake and removal of potentially damaging apoptotic cells is impaired after ethanol treatment. The aim of the present study was to determine whether the presence of apoptotic cells leads to Kupffer cell (KC) production and release of proinflammatory cytokines that have been linked to hepatocyte damage, such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Methods: Kupffer cells were isolated from female rats after an 8-week oral administration of a dextrose control or ethanol-containing fish-oil diet. The isolated KCs were cultured for up to 24 hours in the absence or presence of apoptotic or nonapoptotic hepatoma cells, or lipopolysaccharide. After incubation, media from the cultures were assayed for the presence of TNF-α and IL-6 by immunoassay detection. Also, the expression of these cytokines was measured in KC lysates by a quantitative real-time polymerase chain reaction. Results: Kupffer cells cultured for up to 24 hours in the presence of apoptotic cells produced significantly more TNF-α and IL-6 (80 and 60%, respectively, p<0.05) when the cells were isolated from ethanol-fed animals compared with controls. Additionally, after as early as 4 hours in culture with apoptotic cells, mRNA levels of both cytokines were increased (2-5-fold) in KCs isolated from ethanol-fed animals compared with controls. Conclusions: The presence of apoptotic cells results in the in vitro activation of KCs. Additionally, chronic ethanol administration results in an enhanced responsiveness of KCs to produce proinflammatory cytokines indicated by the increased production of inflammatory mediators from KCs obtained from ethanol-fed animals.

Original languageEnglish (US)
Pages (from-to)122-129
Number of pages8
JournalAlcoholism: Clinical and Experimental Research
Volume31
Issue number1
DOIs
StatePublished - Jan 1 2007

Fingerprint

Kupffer Cells
Rats
Ethanol
Cells
Cytokines
Interleukin-6
Animals
Liver
Fish Oils
Polymerase chain reaction
Nutrition
In Vitro Techniques
Cell culture
Culture Media
Lipopolysaccharides
Immunoassay
Oral Administration
Tumor Necrosis Factor-alpha
Liver Diseases
Real-Time Polymerase Chain Reaction

Keywords

  • Apoptotic Bodies
  • Cytokine
  • Ethanol
  • Kupffer Cells

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Toxicology
  • Psychiatry and Mental health

Cite this

@article{5c4effe2e8f2426aad49c757b10ab08f,
title = "Effect of chronic ethanol administration on the in vitro production of proinflammatory cytokines by rat Kupffer cells in the presence of apoptotic cells",
abstract = "Background: Chronic ethanol consumption can lead to a variety of pathological consequences by as yet undefined mechanisms. Recently, it has been noted that alcohol-associated liver disease is often accompanied by morphological liver changes that include the increased production of apoptotic cells. Additionally, it has been demonstrated that hepatocellular uptake and removal of potentially damaging apoptotic cells is impaired after ethanol treatment. The aim of the present study was to determine whether the presence of apoptotic cells leads to Kupffer cell (KC) production and release of proinflammatory cytokines that have been linked to hepatocyte damage, such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Methods: Kupffer cells were isolated from female rats after an 8-week oral administration of a dextrose control or ethanol-containing fish-oil diet. The isolated KCs were cultured for up to 24 hours in the absence or presence of apoptotic or nonapoptotic hepatoma cells, or lipopolysaccharide. After incubation, media from the cultures were assayed for the presence of TNF-α and IL-6 by immunoassay detection. Also, the expression of these cytokines was measured in KC lysates by a quantitative real-time polymerase chain reaction. Results: Kupffer cells cultured for up to 24 hours in the presence of apoptotic cells produced significantly more TNF-α and IL-6 (80 and 60{\%}, respectively, p<0.05) when the cells were isolated from ethanol-fed animals compared with controls. Additionally, after as early as 4 hours in culture with apoptotic cells, mRNA levels of both cytokines were increased (2-5-fold) in KCs isolated from ethanol-fed animals compared with controls. Conclusions: The presence of apoptotic cells results in the in vitro activation of KCs. Additionally, chronic ethanol administration results in an enhanced responsiveness of KCs to produce proinflammatory cytokines indicated by the increased production of inflammatory mediators from KCs obtained from ethanol-fed animals.",
keywords = "Apoptotic Bodies, Cytokine, Ethanol, Kupffer Cells",
author = "McVicker, {Benita L} and Tuma, {Dean J.} and Kusum Kharbanda and Kubik, {Jacy L.} and Casey, {Carol A}",
year = "2007",
month = "1",
day = "1",
doi = "10.1111/j.1530-0277.2006.00270.x",
language = "English (US)",
volume = "31",
pages = "122--129",
journal = "Alcoholism: Clinical and Experimental Research",
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T1 - Effect of chronic ethanol administration on the in vitro production of proinflammatory cytokines by rat Kupffer cells in the presence of apoptotic cells

AU - McVicker, Benita L

AU - Tuma, Dean J.

AU - Kharbanda, Kusum

AU - Kubik, Jacy L.

AU - Casey, Carol A

PY - 2007/1/1

Y1 - 2007/1/1

N2 - Background: Chronic ethanol consumption can lead to a variety of pathological consequences by as yet undefined mechanisms. Recently, it has been noted that alcohol-associated liver disease is often accompanied by morphological liver changes that include the increased production of apoptotic cells. Additionally, it has been demonstrated that hepatocellular uptake and removal of potentially damaging apoptotic cells is impaired after ethanol treatment. The aim of the present study was to determine whether the presence of apoptotic cells leads to Kupffer cell (KC) production and release of proinflammatory cytokines that have been linked to hepatocyte damage, such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Methods: Kupffer cells were isolated from female rats after an 8-week oral administration of a dextrose control or ethanol-containing fish-oil diet. The isolated KCs were cultured for up to 24 hours in the absence or presence of apoptotic or nonapoptotic hepatoma cells, or lipopolysaccharide. After incubation, media from the cultures were assayed for the presence of TNF-α and IL-6 by immunoassay detection. Also, the expression of these cytokines was measured in KC lysates by a quantitative real-time polymerase chain reaction. Results: Kupffer cells cultured for up to 24 hours in the presence of apoptotic cells produced significantly more TNF-α and IL-6 (80 and 60%, respectively, p<0.05) when the cells were isolated from ethanol-fed animals compared with controls. Additionally, after as early as 4 hours in culture with apoptotic cells, mRNA levels of both cytokines were increased (2-5-fold) in KCs isolated from ethanol-fed animals compared with controls. Conclusions: The presence of apoptotic cells results in the in vitro activation of KCs. Additionally, chronic ethanol administration results in an enhanced responsiveness of KCs to produce proinflammatory cytokines indicated by the increased production of inflammatory mediators from KCs obtained from ethanol-fed animals.

AB - Background: Chronic ethanol consumption can lead to a variety of pathological consequences by as yet undefined mechanisms. Recently, it has been noted that alcohol-associated liver disease is often accompanied by morphological liver changes that include the increased production of apoptotic cells. Additionally, it has been demonstrated that hepatocellular uptake and removal of potentially damaging apoptotic cells is impaired after ethanol treatment. The aim of the present study was to determine whether the presence of apoptotic cells leads to Kupffer cell (KC) production and release of proinflammatory cytokines that have been linked to hepatocyte damage, such as interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Methods: Kupffer cells were isolated from female rats after an 8-week oral administration of a dextrose control or ethanol-containing fish-oil diet. The isolated KCs were cultured for up to 24 hours in the absence or presence of apoptotic or nonapoptotic hepatoma cells, or lipopolysaccharide. After incubation, media from the cultures were assayed for the presence of TNF-α and IL-6 by immunoassay detection. Also, the expression of these cytokines was measured in KC lysates by a quantitative real-time polymerase chain reaction. Results: Kupffer cells cultured for up to 24 hours in the presence of apoptotic cells produced significantly more TNF-α and IL-6 (80 and 60%, respectively, p<0.05) when the cells were isolated from ethanol-fed animals compared with controls. Additionally, after as early as 4 hours in culture with apoptotic cells, mRNA levels of both cytokines were increased (2-5-fold) in KCs isolated from ethanol-fed animals compared with controls. Conclusions: The presence of apoptotic cells results in the in vitro activation of KCs. Additionally, chronic ethanol administration results in an enhanced responsiveness of KCs to produce proinflammatory cytokines indicated by the increased production of inflammatory mediators from KCs obtained from ethanol-fed animals.

KW - Apoptotic Bodies

KW - Cytokine

KW - Ethanol

KW - Kupffer Cells

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U2 - 10.1111/j.1530-0277.2006.00270.x

DO - 10.1111/j.1530-0277.2006.00270.x

M3 - Article

VL - 31

SP - 122

EP - 129

JO - Alcoholism: Clinical and Experimental Research

JF - Alcoholism: Clinical and Experimental Research

SN - 0145-6008

IS - 1

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