Abstract
Kupffer cells contribute to the Important role of the liver defense mechanism through nitric oxide (NO) production. In this study, the effect of chronic ethanol administration on the ability of Kupffer cells to synthesize and release NO was investigated after stimulation with lipopolysaccharide (LPS). Male Wistar rats were chronically fed ethanol for 8 weeks according to the method described by DeCarti and Lieber et al. (J Nutr. 91:331-336, 1967). Kupffer cells were isolated and cultured with LPS (1 μg/ml) for 24 hr. The levels of nitrite and nitrate, metabolites of NO, were determined in the culture medium. NO synthase (NOS) activity in Kupffer cells was determined by the method that measures conversion of [14C]arginine into [14C]citrulline. In control rats, a significant increase of nitrite and nitrate levels in culture medium was observed after LPS treatment. The magnitude of this increase was significantly smaller in chronic ethanol-fed rats. When the activity of NOS was determined, inducible NOS (iNOS) activity was higher than that of constitutive NOS, and LPS administration produced a significant elevation of iNOS activity in both control and chronic ethanol-fed rats. However, the elevation of iNOS activity by LPS stimulation was diminished by chronic ethanol administration. Distribution of iNOS in Kupffer cells as determined by an immunofluorescence method using a laser scanning confocal image system showed a lower expression of iNOS in chronic ethanol-fed rats even in the presence of LPS. These results demonstrate that the excessive production of NO by increased iNOS activity in Kupffer cells is diminished by chronic ethanol administration.
| Original language | English |
|---|---|
| Journal | Alcoholism: Clinical and Experimental Research |
| Volume | 20 |
| Issue number | 1 SUPPL. |
| Publication status | Published - 1996 |
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Keywords
- Laser Scanning Confocal Microscopy
- Lipopolysaccharide
- Nitrate
- Nitric Oxide Synthase
- Nitrite
ASJC Scopus subject areas
- Medicine (miscellaneous)
- Toxicology
Cite this
Effect of chronic ethanol feeding on nitric oxide synthesis by rat Kupffer cells. / Kimura, Hiroyuki; Miura, Soichiro; Higuchi, Hajime; Kurose, Iwao; Tsuzuki, Yoshikazu; Shigematsu, Takeharu; Ebinuma, Hirotoshi; Kato, Shinzo; Ishii, Hiromasa.
In: Alcoholism: Clinical and Experimental Research, Vol. 20, No. 1 SUPPL., 1996.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Effect of chronic ethanol feeding on nitric oxide synthesis by rat Kupffer cells
AU - Kimura, Hiroyuki
AU - Miura, Soichiro
AU - Higuchi, Hajime
AU - Kurose, Iwao
AU - Tsuzuki, Yoshikazu
AU - Shigematsu, Takeharu
AU - Ebinuma, Hirotoshi
AU - Kato, Shinzo
AU - Ishii, Hiromasa
PY - 1996
Y1 - 1996
N2 - Kupffer cells contribute to the Important role of the liver defense mechanism through nitric oxide (NO) production. In this study, the effect of chronic ethanol administration on the ability of Kupffer cells to synthesize and release NO was investigated after stimulation with lipopolysaccharide (LPS). Male Wistar rats were chronically fed ethanol for 8 weeks according to the method described by DeCarti and Lieber et al. (J Nutr. 91:331-336, 1967). Kupffer cells were isolated and cultured with LPS (1 μg/ml) for 24 hr. The levels of nitrite and nitrate, metabolites of NO, were determined in the culture medium. NO synthase (NOS) activity in Kupffer cells was determined by the method that measures conversion of [14C]arginine into [14C]citrulline. In control rats, a significant increase of nitrite and nitrate levels in culture medium was observed after LPS treatment. The magnitude of this increase was significantly smaller in chronic ethanol-fed rats. When the activity of NOS was determined, inducible NOS (iNOS) activity was higher than that of constitutive NOS, and LPS administration produced a significant elevation of iNOS activity in both control and chronic ethanol-fed rats. However, the elevation of iNOS activity by LPS stimulation was diminished by chronic ethanol administration. Distribution of iNOS in Kupffer cells as determined by an immunofluorescence method using a laser scanning confocal image system showed a lower expression of iNOS in chronic ethanol-fed rats even in the presence of LPS. These results demonstrate that the excessive production of NO by increased iNOS activity in Kupffer cells is diminished by chronic ethanol administration.
AB - Kupffer cells contribute to the Important role of the liver defense mechanism through nitric oxide (NO) production. In this study, the effect of chronic ethanol administration on the ability of Kupffer cells to synthesize and release NO was investigated after stimulation with lipopolysaccharide (LPS). Male Wistar rats were chronically fed ethanol for 8 weeks according to the method described by DeCarti and Lieber et al. (J Nutr. 91:331-336, 1967). Kupffer cells were isolated and cultured with LPS (1 μg/ml) for 24 hr. The levels of nitrite and nitrate, metabolites of NO, were determined in the culture medium. NO synthase (NOS) activity in Kupffer cells was determined by the method that measures conversion of [14C]arginine into [14C]citrulline. In control rats, a significant increase of nitrite and nitrate levels in culture medium was observed after LPS treatment. The magnitude of this increase was significantly smaller in chronic ethanol-fed rats. When the activity of NOS was determined, inducible NOS (iNOS) activity was higher than that of constitutive NOS, and LPS administration produced a significant elevation of iNOS activity in both control and chronic ethanol-fed rats. However, the elevation of iNOS activity by LPS stimulation was diminished by chronic ethanol administration. Distribution of iNOS in Kupffer cells as determined by an immunofluorescence method using a laser scanning confocal image system showed a lower expression of iNOS in chronic ethanol-fed rats even in the presence of LPS. These results demonstrate that the excessive production of NO by increased iNOS activity in Kupffer cells is diminished by chronic ethanol administration.
KW - Laser Scanning Confocal Microscopy
KW - Lipopolysaccharide
KW - Nitrate
KW - Nitric Oxide Synthase
KW - Nitrite
UR - http://www.scopus.com/inward/record.url?scp=0029990042&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029990042&partnerID=8YFLogxK
M3 - Article
VL - 20
JO - Alcoholism: Clinical and Experimental Research
JF - Alcoholism: Clinical and Experimental Research
SN - 0145-6008
IS - 1 SUPPL.
ER -