Investigation of the effect of gentisic acid application on the adipogenesis process through transcription factors

The 3T3-L1 precursor adipocyte cell line is derived from Swiss 3T3 mouse embryos and has been widely used as an in vitro model for the transcriptional regulation of adipogenesis. 3T3-L1 precursor adipocytes can differentiate into mature adipocytes under adipogenic conditions. In order to investigate the inhibition of fat cell differentiation, the effects of many natural, synthetic and semi-synthetic substances have been examined, but very few of them have been able to inhibit fat cell differentiation at the desired level. Although there are some studies in the literature showing that benzoic acid inhibits adipogenesis, there are very few studies investigating the effect of gentisic acid, a derivative of benzoic acid, on adipogenesis. In our study, the effect of gentisic acid on the differentiation process and transcriptional factors of 3T3-L1 cells was measured. In our study, 3T3-L1 precursor adipocytes were stimulated with various substances and differentiated into mature adipocytes. In the cell viability test, gentisic acid solutions at concentrations of 50-250 μM, which do not show cytotoxic effect, were applied to differentiated 3T3-L1 cells. Glyceraldehyde-3-phosphate dehydrogenase (GPDH) activity, morphology of the cells was examined by Oil Red O staining and triglyceride content of the cells were measured spectrophotometrically in all undifferentiated and differentiated groups treated with gentisic acid at different concentrations. In addition, the effect of gentisic acid on adipocyte differentiation was examined by looking at the levels of transcription factors (Peroxisome proliferating active receptor-γ (PPAR-γ), CCAAT enhancer binding protein-α (C/EBP-α) and Sterol regulatory element binding protein-1c (SREBP-1c)). Although intracellular triglyceride contents showed a dose-dependent decrease, no statistical difference was found (p>0.05). In the morphological examination, gentisic acid application at increasing concentrations caused a continuous decrease in the size of the cells and intracellular triglyceride storage. Especially high concentrations of gentisic acid treatment caused a decrease in GPDH activity (p<0.05). High concentrations of gentisic acid decreased the levels of all three transcription factors (p<0.05). In the direction of all these results, it is possible to say that gentisic acid inhibits fat cell differentiation.