Malondialdehyde-acetaldehyde-protein adducts increase secretion of chemokines by rat hepatic stellate cells.

Findings obtained from our recent studies have demonstrated that malondialdehyde, a product of lipid peroxidation, and acetaldehyde can react together with proteins in a synergistic manner and form hybrid protein conjugates, which have been designated as malondialdehyde-acetaldehyde (MAA)-protein adducts. These adducts have been detected in livers of ethanol-fed rats and are immunogenic because significant increases in circulating antibody titers against MAA-adducted proteins have been observed in ethanol-fed rats and more recently in human alcoholics. Although immunological factors may tend to perpetuate liver injury, little is known about the direct functional consequences of MAA-adducted proteins on the different cellular populations of the liver. Hepatic stellate cells (HSCs) have been shown to be pivotal in the pathogenesis of fibrosis and in the amplification and self-perpetuation of the inflammatory process. The present study was conducted to determine the effects of MAA-adducted proteins on the function of HSCs. Rat HSCs were exposed to various amounts of MAA-protein adducts and their unmodified controls, and the secretion of two chemokines, monocyte chemoattractant protein (MCP)-1 and macrophage inflammatory protein (MIP)-2, that are involved in the chemotaxis of monocytes/macrophages and neutrophils, respectively, was determined. We observed that bovine serum albumin-MAA induced a dose- and time-dependent increase in the secretion of both of these chemokines. These findings indicate that MAA-adducted proteins may play a role in the modulation of the hepatic inflammatory response and could contribute to the pathogenesis of alcoholic liver disease.

Chronic ethanol ingestion impairs TGF-alpha-stimulated receptor autophosphorylation.

The effects of chronic ethanol feeding on the binding of transforming growth factor-alpha (TGF-alpha) and TGF-alpha-stimulated receptor autophosphorylation were investigated in isolated rat hepatocytes. When hepatocytes were isolated from rats that were fed an ethanol liquid diet for 6-8 weeks, these cells exhibited a marked impairment of TGF-alpha-stimulated autophosphorylation of the receptor that binds this growth factor compared with hepatocytes from the pair-fed controls. This impaired autophosphorylation of receptor tyrosine residues was accompanied by significant decreases in the amount of surface-bound TGF-alpha. Immunoanalysis indicated no changes in receptor number, indicating that decreased receptor content was not responsible for decreased TGF-alpha binding in the hepatocytes from the ethanol-fed rats. In conclusion, chronic ethanol feeding reduced TGF-alpha binding to hepatocytes with a concomitant decrease in the ability of the receptor tyrosine kinase to autophosphorylate its tyrosine residues. These changes were not accompanied by decreased receptor protein content. These defects could lead to altered signal transduction and to impaired reparative and regenerative processes in the liver.

Effects of chronic ethanol administration on the endocytosis of cytokines by rat hepatocytes.

The effects of chronic ethanol administration on the endocytosis of three representative cytokines were investigated in isolated rat hepatocytes. When hepatocytes were isolated from rats that were fed an ethanol liquid diet for 12 to 13 weeks, these cells exhibited a decreased ability to internalize and degrade transforming growth factor-alpha, tumor necrosis factor-alpha and interleukin-6, compared with hepatocytes from the pair-fed controls. This impaired endocytosis of all three cytokines was accompanied by significant decreases in the amount of hepatocyte surface-bound cytokine. Changes in cytokine binding to surface receptors and reduced rates of receptor-cytokine complex internalization into the cells seem to be major contributors to defective endocytosis in hepatocytes from the ethanol-fed rats. Impaired hepatocyte endocytosis could lead to altered steady-state levels of cytokines in the liver and modified physiological responses to cytokines. These changes could affect homeostasis among the various cell types in the liver and could contribute to liver dysfunction and injury.