Cytoprotective Effects of Dietary Fatty Acids on Oxidative Stress in Hepatocytes
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Non-alcoholic fatty liver disease or NAFLD is characterized by the accumulation of fat in the liver, also known as steatosis. This is one of the most common types of liver diseases and can occur in a range of individuals. Once the liver is affected by steatosis, the liver becomes more susceptible to more serious conditions such as non-alcoholic steatohepatitis, cirrhosis, hepatocellular carcinoma, and liver failure. This research focuses on studying the susceptibility of hepatocytes treated with various common dietary fatty acids to subsequent inflammation and stress. Hepatocytes (HepG2 cell line) were treated with oleic, linoleic and palmitic acid at different concentrations and exposed to the pro-oxidant tert-butyl hydroperoxide (t-BHP) to induce oxidative stress. The extent of cytoprotection by fatty acid pre-exposure was determined based on the release of Lactate Dehydrogenase (LDH) into the supernatant from dead/dying cells. After exposure, the cells were assayed for various markers of oxidative stress including membrane integrity, lipid peroxidation, and protein carbonylation. Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) was performed on the carbonylated proteins followed by commassie blue staining for visualization and analysis. In general, samples with higher carbonyl content showed more intact proteins on gels. Our analysis identified four specific bands which were present in samples from cells untreated with t-BHP, as well as fatty acid treated cells. Such bands are most likely proteins that are susceptible to degradation upon carbonylation. Identification of these proteins by mass spectrometry will provide insights into the proteins susceptible to oxidative stress during steatosis, as well as serve as likely biomarkers for complications that arise from steatosis.
Richardson, Thomas C (2010). Cytoprotective Effects of Dietary Fatty Acids on Oxidative Stress in Hepatocytes. Honors and Undergraduate Research. Available electronically from