Gastroenterology

Gastroenterology

Volume 117, Issue 4, October 1999, Pages 942-952
Gastroenterology

Liver, Pancreas, and Biliary Tract
Essential role of tumor necrosis factor α in alcohol-induced liver injury in mice,☆☆

https://doi.org/10.1016/S0016-5085(99)70354-9Get rights and content

Abstract

Background & Aims: Tumor necrosis factor (TNF)-α is associated with increased mortality in alcoholics, but its role in early alcohol-induced liver injury is not fully understood. Recently, it was shown that injury induced by the enteral alcohol delivery model of Tsukamoto and French was reduced by antibodies to TNF-α. To obtain clear evidence for or against the hypothesis that TNF-α is involved, we studied TNF receptor 1 (TNF-R1, p55) or 2 (TNF-R2, p75) knockout mice. Methods: Long-term enteral alcohol delivery was modified for male gene–targeted mice lacking TNF-R1 and TNF-R2. Animals were given a high-fat liquid diet continuously with either ethanol or isocaloric maltose-dextrin as a control for 4 weeks. Results: Ethanol elevated serum levels of alanine aminotransferase nearly 3-fold in wild-type and TNF-R2 knockout mice but not in TNF-R1 knockout mice. Likewise, ethanol caused severe liver injury in wild-type mice (pathology score, 5.5 ± 0.6) and TNF-R2 knockout mice (pathology score, 5.0 ± 0.4), but not in TNF-R1 knockout mice (pathology score, 0.8 ± 0.4; P < 0.001). Conclusions: Long-term ethanol feeding caused liver injury in wild-type and TNF-R2 knockout mice but not in TNF-R1 knockout mice, providing solid evidence in support of the hypothesis that TNF-α plays an important role in the development of early alcohol-induced liver injury via the TNF-R1 pathway. Moreover, the long-term enteral ethanol feeding technique we described for the first time for knockout mice provides a useful new tool for alcohol research.

GASTROENTEROLOGY 1999;117:942-952

Section snippets

Animals

A breeding colony of TNF-R1 knockout mice (p55 −/−) was established at the University of North Carolina at Chapel Hill from breeding pairs kindly donated by Dr. Tak Mak of Amgen Institute (Toronto, Ontario, Canada). Wild-type C57Bl/6J mice and TNF-R2 knockout mice (p75 −/−) were purchased from the Jackson Laboratory (Bar Harbor, ME). Experiments were performed with adult male mice weighing 22-26 g housed in a facility approved by the American Association for Accreditation of Laboratory Animal

Body weights and urine levels of ethanol

Figure 1 shows a typical adult male mouse from this study. All animals survived surgery and gained weight during the subsequent 4 weeks of continuous delivery of the liquid diets with or without ethanol (Table 1). Similar weight gains occurred in wild-type adult mice on chow diet (data not shown), and there were no significant differences in body weights between the groups studied. The reason for the relatively small weight gain observed was that the mice used in this study were adults; similar

Application of continuous enteral alcohol delivery to the mouse

Typical histopathologic findings from livers of wild-type mice fed an ethanol-containing diet indicate that the long-term enteral feeding mouse model is practical. Although surgery in mice is somewhat more difficult than in rats because of their smaller body size, >90% postoperative survival rates can be achieved with care. For optimal survival rates, it was important that the animals be allowed to recover from surgery for 1 week before the liquid diets were initiated. During the recovery

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    Address requests for reprints to: Ronald G. Thurman, Ph.D., Department of Pharmacology, Laboratory of Hepatobiology and Toxicology, CB 7365, Mary Ellen Jones Building, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7365. e-mail: [email protected]; fax: (919) 966-1893.

    ☆☆

    Supported in part by grants from the National Institute on Alcohol Abuse and Alcoholism.

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