Elsevier

Journal of Hepatology

Volume 32, Issue 6, June 2000, Pages 900-910
Journal of Hepatology

Kupffer cell inactivation alleviates ethanol-induced steatosis and CYP2E1 induction but not inflammatory responses in rat liver

https://doi.org/10.1016/S0168-8278(00)80094-XGet rights and content

Abstract

Background/Aims: Gadolinium chloride inactivates Kupffer cells and alleviates alcohol-induced liver lesions. We investigated the mechanism of gadolinium chloride protection after oral ethanol feeding.

Methods: Rats were maintained ethanol-intoxicated for 6 weeks by feeding ethanol in a low-carbohydrate/high-fat liquid diet. Macrophages were inactivated by intravenous administrations of gadolinium chloride. At termination, liver samples and cell lysates obtained from the periportal and perivenous region were analyzed for histopathology, mRNA expression of endotoxin-associated parameters and cytokines and for enzymes involved in oxidative stress.

Results: Ethanol treatment alone caused marked microvesicular/macrovacuolar steatosis and focal inflammation. Gadolinium significantly alleviated pathology, by reducing steatosis but not inflammation. Gadolinium treatment eliminated ED2 immunopositive Kupffer cells, which were larger and more frequent periportally. Ethanol significantly increased the mRNA expression of the endotoxin (LPS) receptor CD14 and the LPS binding protein LBP, but not that of the pro-inflammatory cytokines TNF-α and IL-1β. The mRNA of CD14 was found to be expressed preferentially in the perivenous region, but gadolinium treatment had no significant effect on the expression or the distribution. However, gadolinium significantly moderated the ethanol induction of CYP2E1 and this effect correlated to the degree of steatosis. Ethanol increased glutathione transferase and reduced glutathione peroxidase activity, but these changes persisted after gadolinium treatment.

Conclusions: Our results suggest that gadolinium chloride reduces symptoms of ALD mainly by counteracting steatosis, and that CD14-positive Kupffer cell populations are not involved in gadolinium protection. The strong correlation between pathology and CYP2E1 induction might suggest a steatopathogenic role for this enzyme.

Section snippets

Chronic administration of ethanol

Thirty-one specific pathogen-free male Wistar rats in littermate pairs, weighing 144–188 g, were housed in stainless steel wire-bottom cages and divided in four groups. Two groups received control diet and two ethanol diet. To cause inactivation of Kupffer cells, GdCl3 (Sigma Chemical Co., St. Louis, MO, USA) was injected (10 mg/kg in acidic saline) into the tail vein every third day to one control group and one ethanol group. Controls received acidic saline. The rats were fed for 6 weeks a

Results

All groups of rats gained weight throughout the 6-week period. Although the weight gain of control rats was somewhat higher (170 g±8), it was not significantly different from that of rats treated with ethanol (138 g±8) or with gadolinium only (148 g±4), but the combination of ethanol and gadolinium led to a somewhat reduced weight gain (110g±11, p<0.05). The mean daily ethanol consumption of rats on ethanol diet was 12.6±0.9 g/kg b.wt. and additional gadolinium treatment had no influence

Discussion

This study demonstrates that in the present model GdCl3 alleviates ALD mainly by reducing steatosis, and that there is a concurrent reduction of the ethanolinduced CYP2E1 induction. Treatment with GdCl3 did not significantly reduce inflammation. Thus our results do not support the suggestion that GdCl3 protects primarily via pro-inflammatory pathways, such as CD14-LBP regulated Kupffer cell activation15. The counteracting effect of GdCl3 on steatosis may nevertheless be a crucial protective

Acknowledgements

We are indebted to Dr Päivi Kärkkäinen, Department of Pathology, University Hospital of Helsinki, for valuable guidance in the histopathological evaluation and to Johanna Saarikko for technical assistance. The expert technical assistance of Gunilla Rönnholm throughout this study has been invaluable. This study was supported by grants from the following sources: ACHRI grant 1 R21 AA12031-01 (to KL and MIS), the Oskar Öflund Foundation (to HJ), the Yrjö Jahnsson foundation (to TL) and The Swedish

References (65)

  • C Kilty et al.

    Glutathione S-transferases as biomarkers of organ damage: applications of rodent and canine GST enzyme immunoassays

    Chem Biol Interact

    (1998)
  • D Rüttinger et al.

    In vivo assessment of hepatic alterations following gadolinium chloride-induced Kupffer cell blockade

    J Hepatol

    (1996)
  • T Armbrust et al.

    Functional characterization of two different Kupffer cell populations of normal rat liver

    J Hepatol

    (1996)
  • E Albano et al.

    Role of cytochrome P4502E1-dependent formation of hydroxyethyl free radical in the development of liver damage in rats intragastrically fed with ethanol

    Hepatology

    (1996)
  • M Morimoto et al.

    Modulation of experimental alcohol-induced liver disease by cytochrome P450 2E1 inhibitors

    Hepatology

    (1995)
  • DA Badger et al.

    Gadolinium chloride reduces cytochrome P450: relevance to chemical-induced hepatotoxicity

    Toxicology

    (1997)
  • M Ingelman-Sundberg et al.

    Centrilobular expression of ethanol-inducible cytochrome P-450 (IIE1) in rat liver

    Biochem Biophys Res Commun

    (1988)
  • H Rouach et al.

    Effect of chronic ethanol feeding on lipid peroxidation and protein oxidation in relation to liver pathology

    Hepatology

    (1997)
  • AA Nanji et al.

    Lactobacillus feeding reduces endotoxemia and severity of experimental alcoholic liver disease

    Proc Soc Exp Biol Med

    (1994)
  • HA Järveläinen et al.

    Effect of chronic co-administration of endotoxin and ethanol on rat liver pathology and pro- and anti-inflammatory cytokines

    Hepatology

    (1999)
  • J Hansen et al.

    The role of tumor necrosis factor-α in acute endotoxin-induced hepatotoxicity in ethanol-fed rats

    Hepatology

    (1994)
  • HA Järveläinen et al.

    Alcohol-induced expression of the CD14 endotoxin receptor protein in rat Kupffer cells

    Alcohol Clin Exp Res

    (1997)
  • G Ekstöm et al.

    Rat liver microsomal NADPH-supported oxidase activity and lipid peroxidation dependent on ethanol-inducible cytochrome P450 (P-450 IIE1)

    Biochem Pharmacol

    (1989)
  • DR Koop

    Oxidative and reductive metabolism by cytochrome P-450 2E1

    FASEB J

    (1992)
  • C Fang et al.

    Zonated expression of cytokines in rat liver: effect of chronic ethanol and the cytochrome P450 2E1 inhibitor, chlormethiazole

    Hepatology

    (1998)
  • DR Koop et al.

    Gadolinium chloride blocks alcohol-dependent liver toxicity in rats treated chronically with intragastric alcohol despite the induction of CYP2E1

    Mol Pharmacol

    (1997)
  • H Tsukamoto et al.

    Severe and progressive steatosis and focal necrosis in rat liver induced by continuous intragastric infusion of ethanol and low fat diet

    Hepatology

    (1985)
  • ER Abril et al.

    Binge-type alcohol intake recruits immune cells into rat liver and stimulates Kupffer cell release of cytokines and superoxide prior to any evidence of overt liver injury

  • ER Abril et al.

    Enhanced Kupffer cell function characterizes the recovery period following binge type alcohol intake in rats

  • Z Zhong et al.

    Binge drinking disturbs hepatic microcirculation after transplantation: prevention with free radical scavengers

    J Pharmacol Exp Ther

    (1999)
  • KO Lindros et al.

    A new oral low-carbohydrate alcohol liquid diet producing liver lesions: a preliminary account

    Alcohol Alcohol

    (1998)
  • DL Laskin

    Role of hepatic macrophages in inflammation and tissue injury

  • Cited by (64)

    • Creatine supplementation protects against diet-induced non-alcoholic fatty liver but exacerbates alcoholic fatty liver

      2022, Life Sciences
      Citation Excerpt :

      Studies have reported that alcohol dehydrogenases are responsible for ethanol oxidation in the presence of moderate concentrations of ethanol; in higher concentrations, however, cytochrome P450 (P450) enzymes are more involved [32]. In rats, there is a strong correlation between the degree of steatosis and ethanol activation of CYP2E1 [31,33]. Ethanol fed rats treated with chlormethiazole, a CYP2E1 inhibitor, had blunted ethanol-induced liver fat accumulation and hepatic inflammation [34].

    • Hepatocyte toll-like receptor 4 deficiency protects against alcohol-induced fatty liver disease

      2018, Molecular Metabolism
      Citation Excerpt :

      Kupffer cells, the resident liver macrophages, are a key player in the development of ALD. Deletion of Kupffer cells in rats via gadolinium chloride treatment alleviates alcohol-induced fatty liver disease [44–46]. In addition, mice lacking TLR4 in hematopoietic cells had significantly reduced hepatic triglyceride content [10].

    • Associations between the tumor necrosis factor-α gene and interleukin-10 gene polymorphisms and risk of alcoholic liver disease: A meta-analysis

      2016, Clinics and Research in Hepatology and Gastroenterology
      Citation Excerpt :

      Increased circulating LPS and/or TNF-α was observed in ALD patients, as well as in ethanol-exposed animals [17–21]. Moreover, Kupffer cells inactivation by gadolinium chloride almost completely abolished ethanol-induced liver injury [22–24], while TNF-receptor 1 (TNF-R1) knockout mice were resistant to long-term enteral ethanol feeding compared with the wild type mice [20]. These results provide strong evidence for the hypothesis that TNF-α may play an important role in the development of early alcohol-induced liver injury.

    View all citing articles on Scopus
    View full text