Characterization of miR-100-3p and Other microRNAs in Non-Alcoholic Fatty Liver Disease
Non-Alcoholic Fatty Liver Disease (NAFLD) is a progressive disease characterized by the build-up of fat within the liver, eventually developing liver fibrosis or cirrhosis. NAFLD is the most common form of chronic liver disease in the United States, with a larger incidence in individuals with Type 2 Diabetes or obesity. Previous studies have demonstrated upregulation of the AKR1B10 gene in more advanced stages of NAFLD. Although the gene’s role is currently unknown, recent evidence suggested that certain microRNAs are involved in the regulatory processes of AKR1B10 in advancing stages of NAFLD. By utilizing real-time PCR analysis and western blotting techniques, this project aimed to characterize the roles of miR-100-3p and miR-95-5p in the regulation of AKR1B10 in activated LX-2 cells exposed to glucose or palmitate. Firstly, AKR1B10 expression had a 4.8 fold induction and miR-100-3p had a 6.2 fold decrease in expression in quiescent LX-2 cells when compared to activated LX-2 cells. miR-95-5p had no expression in quiescent cells. AKR1B10 was induced by 15mM and 25mM glucose after 24 hour exposures in activated LX-2 cells. miR-100-3p was significantly induced in activated LX-2 cells treated with 15 or 25mM glucose for 6 hours, but repressed at 24 and 48 hours. Similarly, miR-95-5p was down-regulated by 15mM or 25mM glucose in activated LX-2 cells after 6, 24, or 48 hour exposure. In palmitate treatments, AKR1B10 expression was downregulated by 0.5mM or 1mM palmitate exposures at 6 and 24 hours, whereas miR-100-3p was significantly induced. From these results, there is enough evidence to suggest that miR-100-3p plays a role in regulating AKR1B10 in activated LX-2 cells exposed to glucose or palmitate, while miR-95-5p has no role.