Investigating cytoprotection by the dietary isfolavone Daidzein against palmitic acid cytotoxicity in a human neural cell lineKresal, Martin (Avtor) Gaber, Aljaž (Mentor) Guerra Mollet, Inês (Komentor) type II diabetesdaidzeinpalmitic acidcytotoxicityThe Western diet, high in saturated fats and low in plant-based foods, is associated with metabolic diseases such as type II diabetes. Long-term dietary intervention with a diet high in plant-based foods, rich in phytochemicals has been shown to prevent such diseases. The reason for prevention is not known. We suspected it may be linked to antioxidant effects driven by the presence of phytochemicals in the diet. Phytochemicals are chemicals present in the plant-based part of diet that are not nutrients and do not provide energy or structural components, but they nonetheless influence metabolism. Since isoflavone daidzein is one of the most common dietary phytochemicals and palmitic acid is a dietary saturated fatty acid found in abundance in a Western diet, we investigated whether the dietary isoflavone daidzein is cytoprotective against saturated dietary palmitic acid induced cell damage in a human neural cell line SH-SY5Y. We did that by monitoring the gene expression of mitochondrial transcription factor A encoded by the TFAM gene. Mitochondria both generate reactive oxygen species (ROS) that drive redox sensitive events and respond to ROS-mediated changes in the cellular redox state. Acute stimulation with exogenous oxidants, such as palmitic acid, was shown to increase the concentration of ROS and in turn increase the expression of a key mitochondrial transcription factor TFAM. An increased expression of TFAM has been shown to have cytoprotective effects, since this gene is associated with mitochondrial DNA repair. Therefore, by monitoring the gene expression of TFAM we were able to determine whether daidzein increased expression of this gene and was therefore cytoprotective against palmitic acid induced cell damage. We cultured the cells in conditions with or without palmitic acid, extracted their RNA and used reverse transcription quantitative polymerase chain reaction (RT qPCR) to get the gene expression of the TFAM gene. Quercetin is an isoflavone, which was experimentally proven to increase the TFAM gene expression, so cells cultured with quercetin were used as positive control. Cells cultured without daidzein were used as negative control. As a reference gene, we monitored the gene expression of the B2M gene. The experiment was repeated twice and after both experiments, the statistical analysis showed that the differences in gene expression were not statistically relevant. Due to the high standard deviation, we can only conclude that to reduce the standard deviation between experimental replicates the experiment should be either optimised or repeated. 20252025-05-30 08:50:00Diplomsko delo/naloga169492sl