Year: 2015
Type: dissertation
Abstract: The goal of this diploma thesis was to investigate the effect of S-nitrosoglutathione (GSNO) on adipogenesis of white fat cells using the murine 3T3-L1 cell line. Cells were differentiated according to a standard protocol in the presence of increasing GSNO concentrations and varying times of differentiation. Expression of distinct transcription factors and adipogenic markers were analyzed on mRNA and protein levels using real-time quantitative PCR and Western blot, respectively. Cellular triglycerides were visualized using Nile Red staining and quantified using a commercial kit. Cell number and viability were analyzed by Trypan blue staining.Differentiation of 3T3-L1 cells for 7 days resulted in a pronounced dose-dependent anti-adipogenic effect of GSNO at concentrations of 300 -1000 M. Cell viability was only marginally affected by those relatively high concentrations of GSNO. The anti-adipogenic effect of the thionitrite was also evident as reduced cellular triglyceride content and decreased protein expression of the pro-adipogenic transcription factor sterol regulatory element binding protein 1. Moreover, mRNA levels of peroxisome proliferator-activated receptor and CAAT/ enhancer binding protein were significantly reduced under those conditions. Protein expression of adipocyte-specific targets (hormone sensitive lipase and adipose triglyceride lipase) was significantly downregulated in the presence of GSNO. The anti-adipogenic effect was also observed upon treatment of cells with the nitric oxide donor diethylenetriamine NONOate (100 M). Time course experiments (daily harvest and analysis of cells over a period of 7 days) revealed a prominent anti-proliferative effect evident as decreased cell number and protein content. The molecular mechanism underlying the anti-adipogenic and anti-proliferative effects and the physiological relevance of this finding are not elucidated yet but experiments are ongoing to753i L
Author: Pirker, Teresa Marie
Language: English
Subfield: Physiology (medical)
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W7037239879