Microarray analysis of glucose metabolism-related genes in human umbilical cord blood derived-mesenchymal stem cells treated with everolimus under high glucose conditions

Umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) have attracted attention as a potential cell therapy due to their immunomodulatory properties, paracrine trophic effects, and differentiation capacity that promote tissue repair and functional recovery. However, exposure to diabetic conditions impairs mitochondrial function in UCB-MSCs, leading to reduced stem cell activity and increased cell death, ultimately compromising their therapeutic efficacy. Hyperglycemic stress abnormally activates mTOR signaling, driving dysregulated glucose metabolism and mitochondrial dysfunction. To assess whether everolimus, an mTOR inhibitor, can mitigate these effects, we examined glucose metabolism-related enzyme expression in UCB-MSCs exposed to high glucose with or without everolimus treatment. Microarray profiling targeted 84 glucose metabolism-related genes, including eight housekeeping genes, of which 64 genes were reliably detected. Comparison of high glucose (H) versus high glucose plus everolimus (HE) conditions (two replicates each) revealed limited transcriptional changes, with only a subset of genes showing differential expression between groups. Overall, these results indicate that everolimus does not significantly alter glucose metabolism gene expression in UCB-MSCs. UCB-MSCs were cultured until 70% confluency, then switched to serum-free medium for 24 hours to induce starvation. Cells were assigned to two experimental groups. In the high glucose group (H), UCB-MSCs were exposed to 25 mM D-glucose for 72 h. In the everolimus plus high glucose group (HE), UCB-MSCs were pretreated with 100 nM everolimus for 30 min, followed by the addition of 25 mM D-glucose and incubation for 72 h. Each group included two biological replicates (H1, H2, HE1, HE2).