Liver injury (serum ALT) and steatosis (H&E and oil red staining) were significantly greater in ETOH+Chol and ETOH+Chol+Coil compared to ETOH and ETOH+Coil groups. M1 proinflammatory macrophage markers, MCP-1, Ly6c, TNFα, NOS2 and F4/80, were higher in EtOH+Chol and EtOH+Chol+Coil groups. However,
M2 anti-inflammatory markers, CD163 and Arg-1, were reduced in EtOH+Chol or EtOH+Chol+Coil fed groups. Furthermore, the inflammatory markers IL-6 and ICAM-1 were more elevated in EtOH+Chol and EtOH+Chol+Coil groups. Additionally, fribrosis examined by Sirius red staining and fibrotic markers, collagen, αSMA, osteopontin, desmin and hydroxyproline levels were highly increased in EtOH+Chol and EtOH+Chol+Coil groups. Alcohol GDC-0068 molecular weight binge drinking caused more injury (serum ALT and H&E) and inflammation Trametinib in HC than in LD pretreated mice despite similar degree of steatosis. Conclusion: cholesterol intake has a synergic effect with alcohol aggravating liver injury and progression to alcoholic hepatitis, pointing that targeting cholesterol may be a valuable approach for future therapeutic interventions in ALD. Disclosures: The following people have nothing to disclose: Laura Conde de la Rosa, Carmen Garcia-Ruiz, Jose Fernandez-Checa
Alcoholic steatohepatitis (ASH) and nonalcoholic steatohepati-tis (NASH) are the most frequent conditions leading to medchemexpress elevated liver enzymes and liver cirrhosis, respectively, in the Western world. However, despite strong epidemiological evidence for combined effects on the progression of liver injury, the mutual interaction of the pathophysiological mechanisms is incompletely understood. The aim of this study was to establish an in vitro model for joint effects of alcohol and lipids on hepatic steatosis and inflammation. Methods and Results: Initially, we established the dose
range in which neither alcohol nor incubation with the free fatty acid oleate affected viability or mito-chondrial activity in primary human hepatocytes (PHH). Subsequently, we assessed the combined effect of alcohol (1%ndash;2%) and oleate (0.2mM) on hepatocellular lipid accumulation in PHH. Under these conditions, alcohol significantly enhanced oleate induced expression of genes regulating lipogenesis (FASN, SCD-1) and lipid peroxidation (CPT-1) as well as cellular triglyceride content and free fatty acid (FFA) levels, while alcohol alone had only a minimal effect. Analysis of heme oxy-genase-1 (HMOX-1) expression and malondialdehyde levels revealed that the combination of alcohol and oleate caused significantly higher oxidative stress and lipid peroxidation than either of the two substances alone. Further, we observed a syn-ergistic effect of alcohol and FFA on JNK-activation and pro-inflammatory (IL-8 and ICAM-1) gene expression.