Hepatic TLR4 Signaling in Obese NAFLD.
AUTHORS
- PMID: 26113297 [PubMed].
ABSTRACT
Nonalcoholic fatty liver disease (NAFLD) 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 (NAFL) 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 to NAFL as was IRF3 in the myeloid differentiation factor 88- (Myd88-) independent signaling pathway. In HepaRG cells, NF-ҡB nuclear translocation and functional activity increased following treatment with the fatty acid, palmitate, and following exposure to LPS, when compared to hepatocytes stimulated with a lipogenic treatment that induced de novo lipogenesis. Palmitate- and LPS-induction of NF-ҡB activity was partially attenuated by chemical- or siRNA-mediated inhibition of TLR4. Expression of TLR4 and its downstream mediators was up-regulated with palmitate and LPS. Similar results were observed using primary human hepatocytes from a lean donor. Interestingly, NF-ҡB activity assays showed obese donor hepatocytes were resistant to chemical TLR4 inhibition.
Nonalcoholic fatty liver disease (NAFLD) 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 (NAFL) 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 to NAFL as was IRF3 in the myeloid differentiation factor 88- (Myd88-) independent signaling pathway. In HepaRG cells, NF-ҡB nuclear translocation and functional activity increased following treatment with the fatty acid, palmitate, and following exposure to LPS, when compared to hepatocytes stimulated with a lipogenic treatment that induced de novo lipogenesis. Palmitate- and LPS-induction of NF-ҡB activity was partially attenuated by chemical- or siRNA-mediated inhibition of TLR4. Expression of TLR4 and its downstream mediators was up-regulated with palmitate and LPS. Similar results were observed using primary human hepatocytes from a lean donor. Interestingly, NF-ҡB activity assays showed obese donor hepatocytes were resistant to chemical TLR4 inhibition.
Tags: Alumni Publications 2015