Summary
Chronic consumption of ethanol induces hepatic steatosis and inflammation, which can eventually lead to more severe liver injury, characterized by fibrosis and cirrhosis. Recruitment of neutrophils to the liver, as well as activation of Kupffer cells, mediates the inflammatory responses observed after chronic ethanol exposure. Kupffer cells, the resident macrophages of the liver, are critical to the onset of ethanol-induced liver injury. Activation of Kupffer cells leads to an increased production of proinflammatory cytokines, such as tumor necrosis factor-α and also reactive oxygen species, a process mediated in part by changes in lipopolysaccharide-induced TLR4-dependent signal transduction. The isolation and culture of Kupffer cells is an important technique with which one can elucidate the mechanisms that contribute to alcoholic liver injury.
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References
1. Adachi, Y., Bradford, B. U., Gao, W., Bojes, H. K., and Thurman, R. G. (1994) Inactivation of Kupffer cells prevents early alcohol-induced liver injury. Hepatology 20, 453–460.
2. Aldred, A., and Nagy, L. E. (1999) Ethanol dissociates hormone-stimulated camp production from inhibition of TNFα production in rat Kupffer cells. Am. J. Physiol. Gastrointest. Liver Physiol. 276, G98–G106.
3. Nagy, L. E. (2003) New insights into the role of the innate immune response in the development of alcoholic liver disease. Exp. Biol. Med. 228, 882–890.
4. Thurman RG II (1998). Alcoholic liver injury involves activation of Kupffer cells by endotoxin. Am. J. Physiol. 275, G605–G611.
5. Thakur, V., Pritchard, M. T., McMullen, M. R., and Nagy, L. E. (2006) Adiponectin normalizes LPS-stimulated TNFα production by rat Kupffer cells after chronic ethanol feeding. Am. J. Physiol. Gastrointest. Liver Physiol. 290, G998–G1007.
6. Thakur, V. Pritchard, M. T., McMullen, M. R., Wang, Q., and Nagy, L. E. (2006) Chronic ethanol feeding increases activation of NADPH oxidase by lipopolysaccharide in rat Kupffer cells: role of increased reactive oxygen in LPS-stimulated ERK1/2 activation and TNFα production. J. Leukoc. Biol. 79, 1348–1356.
7. Lieber, C. M., and DeCarli, L. M. (1982) The feeding of alcohol in liquid diets: two decades of application and 1982 update. Alcohol. Clin. Exp. Res. 6, 523–531.
Friedman, S. L., and Roll, F. J. Isolation and culture of hepatic lipocytes, Kupffer cells, and sinusoidal endothelial cells by density gradient centrifugation with stractan. Anal. Biochem. 161, 207–218.
9. Knook, D. L., Blansjaar, N., and Sleyster, E. C. (1977) Isolation and characterization of Kupffer and endothelial cells from the rat liver. Exp. Cell Res. 109, 317–329.
10. Munthe-Kaas, A. C., Berg, T., Seglen, P. O., and Seljelid, R. (1975) Mass Isolation and culture of rat Kupffer cells. J. Exp. Med. 141, 1–10.
Acknowledgements
The work in this chapter was supported by National Institute on Alcohol Abuse and Alcoholism Grant no. AA-013868 and AA-011975 (to L.E.N.) and AA-015833 (to M.T.P.).
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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McMullen, M.R., Pritchard, M.T., Nagy, L.E. (2008). Isolation of Kupffer Cells from Rats Fed Chronic Ethanol. In: Nagy, L.E. (eds) Alcohol. Methods in Molecular Biology™, vol 447. Humana Press. https://doi.org/10.1007/978-1-59745-242-7_15
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DOI: https://doi.org/10.1007/978-1-59745-242-7_15
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