Myeloid Mixed Lineage Kinase 3 Contributes to Chronic Ethanol-Induced Inflammation and Hepatocyte Injury in Mice

• Published in: Gene expression 2016, Vol.17 (1), p.61-77
• Main Authors: McCullough, Rebecca L., Saikia, Paramananda, Pollard, Katherine A., McMullen, Megan R., Nagy, Laura E., Roychowdhury, Sanjoy
• Format: Article
• Language: English
• Subjects: Alcoholic Liver Disease (ald); Animals; Apoptosis; Apoptosis - drug effects; Bone marrow; Cell activation; Cell death; Cytokeratin; Cytokines; Diet; Ethanol; Ethanol - adverse effects; Female; Females; Hepatocytes - drug effects; Hepatocytes - metabolism; Inflammation; Inflammation - chemically induced; Inflammation - metabolism; Injuries; Kinases; Kupffer Cells; Kupffer Cells - drug effects; Kupffer Cells - metabolism; Lipopolysaccharides - pharmacology; Liver; Liver - drug effects; Liver - metabolism; Liver diseases; Liver Diseases, Alcoholic - etiology; Liver Diseases, Alcoholic - metabolism; Macrophages; MAP kinase; MAP kinase kinase; MAP Kinase Kinase 4 - metabolism; MAP Kinase Kinase Kinases - metabolism; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase Kinase Kinase 11; Myeloid cells; Necroptosis; Phosphorylation; Phosphorylation - drug effects; Rats; Rats, Wistar; Steatosis; Toll-Like Receptor 4 (tlr4); Transplants & implants; Tumor necrosis factor-TNF
• ISSN: 1052-2166; 1555-3884
• DOI: 10.3727/105221616X691730

Proinflammatory activity of hepatic macrophages plays a key role during progression of alcoholic liver disease (ALD). Since mixed lineage kinase 3 (MLK3)-dependent phosphorylation of JNK is involved in the activation of macrophages, we tested the hypothesis that myeloid MLK3 contributes to chronic ethanol-induced inflammatory responses in liver, leading to hepatocyte injury and cell death. Primary cultures of Kupffer cells, as well in vivo chronic ethanol feeding, were used to interrogate the role of MLK3 in the progression of liver injury. Phosphorylation of MLK3 was increased in primary cultures of Kupffer cells isolated from ethanol-fed rats compared to cells from pair-fed rats. Kupffer cells from ethanol-fed rats were more sensitive to LPS-stimulated cytokine production; this sensitization was normalized by pharmacological inhibition of MLK3. Chronic ethanol feeding to mice increased MLK3 phosphorylation robustly in F4/80+ Kupffer cells, as well as in isolated nonparenchymal cells. MLK3−/− mice were protected from chronic ethanol-induced phosphorylation of MLK3 and JNK, as well as multiple indicators of liver injury, including increased ALT/AST, inflammatory cytokines, and induction of RIP3. However, ethanol-induced steatosis and hepatocyte apoptosis were not affected by MLK3. Finally, chimeric mice lacking MLK3 only in myeloid cells were also protected from chronic ethanol-induced phosphorylation of JNK, expression of inflammatory cytokines, and increased ALT/AST. MLK3 expression in myeloid cells contributes to phosphorylation of JNK, increased cytokine production, and hepatocyte injury in response to chronic ethanol. Our data suggest that myeloid MLK3 could be targeted for developing potential therapeutic strategies to suppress liver injury in ALD patients.