CD18/ICAM-1-dependent nitric oxide production of Kupffer cells as a cause of mitochondrial dysfunction in hepatoma cells

Influence of chronic alcohol feeding

Iwao Kurose, Hajime Higuchi, Naoyuki Watanabe, Soichiro Miura, Kengo Tomita, Yoshikazu Yonei, Masaaki Takaishi, Shigeyuki Zeki, Tetsuya Nakamura, Hidetsugu Saito, Shinzo Kato, Hiromasa Ishii

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The present study was designed to monitor the process for hepatoma cell injury induced by Kupffer cells. The non-activated Kupffer cells isolated from male Wistar rats reduced the mitochondrial membrane potential in the cocultured AH70 cells, which was indicated by the decreased rhodamine 123 (Rh123) fluorescence. Increased level of nitrite and nitrate in the medium and induction of iNOS in Kupffer cells were observed after coculture with AH70 cells. Incubation with either N(G)-monomethyl-L-arginine or aminoguanidine attenuated the increased nitric oxide (NO) production of Kupffer cells and the decreased Rh123 fluorescence of AH70 cells. Fluo-3, a calcium-sensitive probe, fluorescence in Kupffer cells increased after coculture with AH70 cells. Addition of TMB-8, a calcium inhibitor, or monoclonal antibody directed against ICAM-1 or CD18 prevented the increases in fluo-3 fluorescence and NO production of Kupffer cells and Kupffer cell- induced mitochondrial dysfunction in AH70 cells, suggesting the involvement of calcium mobilization and CD18/ICAM-1. It is therefore suggested that the Kupffer cell-mediated mitochondrial dysfunction of hepatoma cells largely depends on NO production by iNOS, and that the NO production by Kupffer cells is triggered by CD18/ICAM-1-dependent interaction with hepatoma cells and subsequent calcium mobilization. In other series of experiments, male Wistar rats fed ethanol for 4 weeks were used. The NO production and calcium mobilization of Kupffer cells and reduction of the mitochondrial membrane potential in cocultured hepatoma cells were diminished in the case of Kupffer cells isolated from chronically ethanol-fed rats, while CD18 and ICAM-1 expression was still observed. Thus, the present study further suggests that NO-dependent anti-hepatoma cell activity of Kupffer cells is suppressed in chronically ethanol-fed animals.

Original languageEnglish
Pages (from-to)229-239
Number of pages11
JournalFree Radical Biology and Medicine
Volume22
Issue number1-2
DOIs
Publication statusPublished - 1996

Fingerprint

Kupffer Cells
Intercellular Adhesion Molecule-1
Hepatocellular Carcinoma
Nitric Oxide
Alcohols
Calcium
Fluorescence
Rhodamine 123
Rats
Ethanol
Mitochondrial Membrane Potential
Coculture Techniques
Wistar Rats
Membranes
Nitrites
Nitrates
Arginine
Animals
Monoclonal Antibodies

Keywords

  • A23187
  • Aminoguanidine
  • Fluo-3
  • Laser scanning confocal microscope
  • N(G)-monomethyl-L-arginine (L-NMMA)
  • Rhodamine 123

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Medicine(all)
  • Toxicology

Cite this

CD18/ICAM-1-dependent nitric oxide production of Kupffer cells as a cause of mitochondrial dysfunction in hepatoma cells : Influence of chronic alcohol feeding. / Kurose, Iwao; Higuchi, Hajime; Watanabe, Naoyuki; Miura, Soichiro; Tomita, Kengo; Yonei, Yoshikazu; Takaishi, Masaaki; Zeki, Shigeyuki; Nakamura, Tetsuya; Saito, Hidetsugu; Kato, Shinzo; Ishii, Hiromasa.

In: Free Radical Biology and Medicine, Vol. 22, No. 1-2, 1996, p. 229-239.

Research output: Contribution to journalArticle

Kurose, Iwao ; Higuchi, Hajime ; Watanabe, Naoyuki ; Miura, Soichiro ; Tomita, Kengo ; Yonei, Yoshikazu ; Takaishi, Masaaki ; Zeki, Shigeyuki ; Nakamura, Tetsuya ; Saito, Hidetsugu ; Kato, Shinzo ; Ishii, Hiromasa. / CD18/ICAM-1-dependent nitric oxide production of Kupffer cells as a cause of mitochondrial dysfunction in hepatoma cells : Influence of chronic alcohol feeding. In: Free Radical Biology and Medicine. 1996 ; Vol. 22, No. 1-2. pp. 229-239.
@article{b22d08eb5d064ffda8dfa67600537273,
title = "CD18/ICAM-1-dependent nitric oxide production of Kupffer cells as a cause of mitochondrial dysfunction in hepatoma cells: Influence of chronic alcohol feeding",
abstract = "The present study was designed to monitor the process for hepatoma cell injury induced by Kupffer cells. The non-activated Kupffer cells isolated from male Wistar rats reduced the mitochondrial membrane potential in the cocultured AH70 cells, which was indicated by the decreased rhodamine 123 (Rh123) fluorescence. Increased level of nitrite and nitrate in the medium and induction of iNOS in Kupffer cells were observed after coculture with AH70 cells. Incubation with either N(G)-monomethyl-L-arginine or aminoguanidine attenuated the increased nitric oxide (NO) production of Kupffer cells and the decreased Rh123 fluorescence of AH70 cells. Fluo-3, a calcium-sensitive probe, fluorescence in Kupffer cells increased after coculture with AH70 cells. Addition of TMB-8, a calcium inhibitor, or monoclonal antibody directed against ICAM-1 or CD18 prevented the increases in fluo-3 fluorescence and NO production of Kupffer cells and Kupffer cell- induced mitochondrial dysfunction in AH70 cells, suggesting the involvement of calcium mobilization and CD18/ICAM-1. It is therefore suggested that the Kupffer cell-mediated mitochondrial dysfunction of hepatoma cells largely depends on NO production by iNOS, and that the NO production by Kupffer cells is triggered by CD18/ICAM-1-dependent interaction with hepatoma cells and subsequent calcium mobilization. In other series of experiments, male Wistar rats fed ethanol for 4 weeks were used. The NO production and calcium mobilization of Kupffer cells and reduction of the mitochondrial membrane potential in cocultured hepatoma cells were diminished in the case of Kupffer cells isolated from chronically ethanol-fed rats, while CD18 and ICAM-1 expression was still observed. Thus, the present study further suggests that NO-dependent anti-hepatoma cell activity of Kupffer cells is suppressed in chronically ethanol-fed animals.",
keywords = "A23187, Aminoguanidine, Fluo-3, Laser scanning confocal microscope, N(G)-monomethyl-L-arginine (L-NMMA), Rhodamine 123",
author = "Iwao Kurose and Hajime Higuchi and Naoyuki Watanabe and Soichiro Miura and Kengo Tomita and Yoshikazu Yonei and Masaaki Takaishi and Shigeyuki Zeki and Tetsuya Nakamura and Hidetsugu Saito and Shinzo Kato and Hiromasa Ishii",
year = "1996",
doi = "10.1016/S0891-5849(96)00332-2",
language = "English",
volume = "22",
pages = "229--239",
journal = "Free Radical Biology and Medicine",
issn = "0891-5849",
publisher = "Elsevier Inc.",
number = "1-2",

}

TY - JOUR

T1 - CD18/ICAM-1-dependent nitric oxide production of Kupffer cells as a cause of mitochondrial dysfunction in hepatoma cells

T2 - Influence of chronic alcohol feeding

AU - Kurose, Iwao

AU - Higuchi, Hajime

AU - Watanabe, Naoyuki

AU - Miura, Soichiro

AU - Tomita, Kengo

AU - Yonei, Yoshikazu

AU - Takaishi, Masaaki

AU - Zeki, Shigeyuki

AU - Nakamura, Tetsuya

AU - Saito, Hidetsugu

AU - Kato, Shinzo

AU - Ishii, Hiromasa

PY - 1996

Y1 - 1996

N2 - The present study was designed to monitor the process for hepatoma cell injury induced by Kupffer cells. The non-activated Kupffer cells isolated from male Wistar rats reduced the mitochondrial membrane potential in the cocultured AH70 cells, which was indicated by the decreased rhodamine 123 (Rh123) fluorescence. Increased level of nitrite and nitrate in the medium and induction of iNOS in Kupffer cells were observed after coculture with AH70 cells. Incubation with either N(G)-monomethyl-L-arginine or aminoguanidine attenuated the increased nitric oxide (NO) production of Kupffer cells and the decreased Rh123 fluorescence of AH70 cells. Fluo-3, a calcium-sensitive probe, fluorescence in Kupffer cells increased after coculture with AH70 cells. Addition of TMB-8, a calcium inhibitor, or monoclonal antibody directed against ICAM-1 or CD18 prevented the increases in fluo-3 fluorescence and NO production of Kupffer cells and Kupffer cell- induced mitochondrial dysfunction in AH70 cells, suggesting the involvement of calcium mobilization and CD18/ICAM-1. It is therefore suggested that the Kupffer cell-mediated mitochondrial dysfunction of hepatoma cells largely depends on NO production by iNOS, and that the NO production by Kupffer cells is triggered by CD18/ICAM-1-dependent interaction with hepatoma cells and subsequent calcium mobilization. In other series of experiments, male Wistar rats fed ethanol for 4 weeks were used. The NO production and calcium mobilization of Kupffer cells and reduction of the mitochondrial membrane potential in cocultured hepatoma cells were diminished in the case of Kupffer cells isolated from chronically ethanol-fed rats, while CD18 and ICAM-1 expression was still observed. Thus, the present study further suggests that NO-dependent anti-hepatoma cell activity of Kupffer cells is suppressed in chronically ethanol-fed animals.

AB - The present study was designed to monitor the process for hepatoma cell injury induced by Kupffer cells. The non-activated Kupffer cells isolated from male Wistar rats reduced the mitochondrial membrane potential in the cocultured AH70 cells, which was indicated by the decreased rhodamine 123 (Rh123) fluorescence. Increased level of nitrite and nitrate in the medium and induction of iNOS in Kupffer cells were observed after coculture with AH70 cells. Incubation with either N(G)-monomethyl-L-arginine or aminoguanidine attenuated the increased nitric oxide (NO) production of Kupffer cells and the decreased Rh123 fluorescence of AH70 cells. Fluo-3, a calcium-sensitive probe, fluorescence in Kupffer cells increased after coculture with AH70 cells. Addition of TMB-8, a calcium inhibitor, or monoclonal antibody directed against ICAM-1 or CD18 prevented the increases in fluo-3 fluorescence and NO production of Kupffer cells and Kupffer cell- induced mitochondrial dysfunction in AH70 cells, suggesting the involvement of calcium mobilization and CD18/ICAM-1. It is therefore suggested that the Kupffer cell-mediated mitochondrial dysfunction of hepatoma cells largely depends on NO production by iNOS, and that the NO production by Kupffer cells is triggered by CD18/ICAM-1-dependent interaction with hepatoma cells and subsequent calcium mobilization. In other series of experiments, male Wistar rats fed ethanol for 4 weeks were used. The NO production and calcium mobilization of Kupffer cells and reduction of the mitochondrial membrane potential in cocultured hepatoma cells were diminished in the case of Kupffer cells isolated from chronically ethanol-fed rats, while CD18 and ICAM-1 expression was still observed. Thus, the present study further suggests that NO-dependent anti-hepatoma cell activity of Kupffer cells is suppressed in chronically ethanol-fed animals.

KW - A23187

KW - Aminoguanidine

KW - Fluo-3

KW - Laser scanning confocal microscope

KW - N(G)-monomethyl-L-arginine (L-NMMA)

KW - Rhodamine 123

UR - http://www.scopus.com/inward/record.url?scp=0030636529&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030636529&partnerID=8YFLogxK

U2 - 10.1016/S0891-5849(96)00332-2

DO - 10.1016/S0891-5849(96)00332-2

M3 - Article

VL - 22

SP - 229

EP - 239

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - 1-2

ER -