title: Hepatic TLR4 signaling in obese NAFLD.
abstract: Nonalcoholic fatty liver disease occurs frequently in the setting of metabolic syndrome, but the factors leading to nonalcoholic steatohepatitis (NASH) are not fully understood. This study investigated Toll-like receptor 4 (TLR4) signaling in human liver with the goal of delineating whether activation of this pathway segregates those with nonalcoholic fatty liver from those with NASH. Experiments were performed using liver biopsy tissue obtained from class III obese subjects undergoing bariatric surgery, and extended to an immortalized human hepatocyte HepaRG cell line and primary human hepatocytes. The bacterial endotoxin lipopolysaccharide (LPS) and total free fatty acid levels were significantly increased in plasma of NASH patients. TLR4 mRNA levels were significantly increased in subjects with NASH compared with NAFL as was interferon regulatory factor (IRF) 3 in the myeloid differentiation factor 88-independent signaling pathway. In HepaRG cells, nuclear factor-κB (NF-κB) nuclear translocation and functional activity increased following treatment with the fatty acid, palmitate, and following exposure to LPS compared with hepatocytes stimulated with a lipogenic treatment that induced de novo lipogenesis. Palmitate and LPS induction of NF-κB activity was partially attenuated by chemical- or small-interfering RNA-mediated inhibition of TLR4. Expression of TLR4 and its downstream mediators was upregulated with palmitate and LPS. Similar results were observed using primary human hepatocytes from a lean donor. Interestingly, NF-κB activity assays showed obese donor hepatocytes were resistant to chemical TLR4 inhibition. In conclusion, TLR4 expression is upregulated in a large cohort of NASH patients, compared with those with NAFL, and this occurs within the setting of increased LPS and fatty acids.
authors.affiliation: Division of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, Tennessee;
authors.forename: Torfay
authors.initials: T
authors.lastname: Sharifnia
authors.affiliation: Department of Surgery, Vanderbilt University Medical Center, Nashville, Tennessee; and.
authors.forename: Joseph
authors.initials: J
authors.lastname: Antoun
authors.affiliation: Division of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, Tennessee;
authors.forename: Thomas G C
authors.initials: TG
authors.lastname: Verriere
authors.affiliation: Division of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, Tennessee;
authors.forename: Giovanni
authors.initials: G
authors.lastname: Suarez
authors.affiliation: Division of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, Tennessee;
authors.forename: Julia
authors.initials: J
authors.lastname: Wattacheril
authors.affiliation: Division of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, Tennessee; Veterans Affairs Tennessee Valley Healthcare System, Nashville, Tennessee.
authors.forename: Keith T
authors.initials: KT
authors.lastname: Wilson
authors.affiliation: Division of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Vanderbilt University Medical Center, Nashville, Tennessee;
authors.forename: Richard M
authors.initials: RM
authors.lastname: Peek
authors.affiliation: Department of Surgery, Vanderbilt University Medical Center, Nashville, Tennessee; and.
authors.forename: Naji N
authors.initials: NN
authors.lastname: Abumrad
authors.affiliation: Department of Surgery, Vanderbilt University Medical Center, Nashville, Tennessee; and robb.flynn@vanderbilt.edu.
authors.forename: Charles R
authors.initials: CR
authors.lastname: Flynn
mesh_terms: D000328:Adult; D002460:Cell Line; D002478:Cells, Cultured; D005260:Female; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D050838:Interferon Regulatory Factor-3 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000097:blood / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D053594:Myeloid Differentiation Factor 88 / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D065626:Non-alcoholic Fatty Liver Disease / Q000150:complications / Q000378:metabolism*; D009765:Obesity / Q000150:complications / Q000378:metabolism*; D019308:Palmitic Acid / Q000097:blood / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*
chemical_list: C494232:IRF3 protein, human; D050838:Interferon Regulatory Factor-3; D008070:Lipopolysaccharides; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; D012333:RNA, Messenger; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D019308:Palmitic Acid
keywords: Toll-like receptor 4; lipopolysaccharide; nonalcoholic fatty liver; nonalcoholic fatty liver disease; nonalcoholic steatohepatitis; nuclear factor-κB; palmitate