the In Vitro and In Silico Analysis of the Potential Effects of Kaempferol on Cell Viability and Calcium-Mediated Apoptotic Pathways in C6 Glioma Cells

Abstract

This study aimed to investigate the effects of kaempferol on cell viability, intracellular calcium signaling, and apoptosis-related possible molecular mechanisms in C6 glioma cells. C6 rat glioma cells were cultured under appropriate cell culture conditions and treated with 25, 50, and 100 µM concentrations of kaempferol for 24 hours. Cell viability was evaluated using the XTT assay. Changes in intracellular calcium levels were analyzed by fluorescent calcium imaging using Fluo-4 AM fluorescent dye. To evaluate the potential effects of kaempferol on calcium-mediated apoptotic mechanisms, a protein–protein interaction (PPI) network including BAX, VDAC1, CYCS, CASP9, and CASP3 proteins was constructed using the STRING database. In addition, KEGG pathway enrichment analysis was performed for the selected gene/protein set. Statistical analyses were conducted using one-way ANOVA followed by Tukey’s post hoc test, and p<0.05 was considered statistically significant. The findings demonstrated that kaempferol treatment significantly reduced the viability of C6 glioma cells and increased intracellular calcium signaling. In conclusion, kaempferol may exert antiproliferative effects in C6 glioma cells through calcium-mediated apoptotic mechanisms. However, further molecular and functional studies are needed to confirm the underlying mechanism more comprehensively.