Small Molecule Inhibitors of Inflammation Prevent Hepatic Steatosis in High Fat Diet-Fed Male C57BL/6J Mice

Presentation Number: MON 510
Date of Presentation: April 3rd, 2017

Ashley Patton*1, Caroline Wilson1, Doug Goetz1, Stephen Bergmeier1, Frank L Schwartz2 and Kelly Dawn McCall3
1Ohio University, 2Ohio University Heritage College of Osteopathic Medicine, 3Ohio University Heritage College of Osteopathic Medicine, Athens, OH


Nonalcoholic fatty liver disease (NAFLD) is the hepatic manifestation of both metabolic and inflammatory diseases, and it has become pervasive worldwide. Inflammation, including inflammation resulting from free fatty acid (FFA) activation of toll-like receptor (TLR) signaling, has been suggested to be an essential component of the pathophysiology of NAFLD. High fat diets (HFDs) promote an increased uptake and storage of FFAs and triglycerides (TGs) in hepatocytes, which initiates steatosis and induces lipotoxicity and inflammation. The objectives of this study were to evaluate the efficacy of novel small molecule inhibitors of inflammation developed in our laboratory, including a TLR inhibitor [phenylmethimazole (C10)], to delay and/or prevent HFD-induced steatosis in a C57BL/6J mouse model of high fat diet (HFD)-induced NAFLD. The mice were fed a HFD (60 % fat, 20% protein, 70% carbohydrate) and divided into 4 groups (N=5 for each group): sham injection (stress control), DMSO (vehicle control), phenylmethimazole (C10), and COB-214. Each compound was administered once daily at a dosage of 1mg/kg for 6 weeks. Histological examination of liver sections from mice in this study using hematoxylin and eosin (H&E) and Oil Red O staining revealed less hepatic lipid accumulation in HFD-fed mice treated with C10 and COB-214 when compared to HFD-fed sham and DMSO groups. Hepatic triglyceride quantification analysis revealed that liver tissues of HFD-fed mice treated with C10 and COB-214 had less triglyceride than mice in the HFD-fed sham and DMSO control groups. COB-214 had the greatest inhibitory activity. Inhibition of TLR4 signaling in palmitate-treated HepG2 cells was measured by reporter assay and real-time PCR. Transcription of pro-inflammatory cytokines was down-regulated in C10 and COB-214 treated groups when compared to Palmitate and Palmitate + DMSO treated groups. Hepatic lipogenesis was decreased by measurement of acetyl-CoA carboxylase (Acc) mRNA levels in C10 and COB-214 treated groups. Future directions for this study include an extended study to determine the mechanisms by which COB-214 prevents the development of nonalcoholic fatty liver disease at several additional time points (4 and 16 weeks). At the conclusion of this study, we hope to have established the role of a novel class of small molecule inhibitors of inflammation in the treatment/prevention of NAFLD.


Nothing to Disclose: AP, CW, DG, SB, FLS, KDM