Abstract:
Background Aging is the inevitable result of the comprehensive action of physiological or pathological processes. Research on how to delays aging, especially pathological aging, can provide new ideas for the treatment of many diseases. The establishment of aging cell model is the cornerstone of aging related research.
Objective To explore the effect of Dulbecco's Modified Eagle Medium (DMEM) with different concentrations of glucose on skin fibroblast aging.
Methods Fibroblasts were cultured in DMEM medium containing different concentrations of glucose (5.5 mmol/L, 15 mmol/L, 25 mmol/L, 35 mmol/L and 45 mmol/L). Cell proliferation and migration were detected by CCK-8 test and cell scratch assay, respectively. β-galactosidase staining was employed to determine the percentage of senescent cell. PCR was used to detect the expression levels of p53, p21 and p16 in each group to elucidate the underlying mechanism.
Results Compared with the 5.5 mmol/L group, the proliferation and migration of fibroblasts in the 15 mmol/L and the 25 mmol/L groups increased, while significantly decreased in the 35 mmol/L and the 45 mmol/L groups; the proportion of SA-β-gal positive cells increased along with the glucose concentration. Meanwhile, the expressions of p53, p21 and p16 were also positively correlated with glucose concentrations.
Conclusion Excessively high concentration of glucose (≥35 mmol/L) inhibits the biological function of cells and induces cell senescence. The pathogenesis is associated with increased expression of p16 via upregulation of p53/ p21 pathway.