RESUMEN
BACKGROUND:Although vitamin C has an anti-oxidation role and can promote cell proliferation, there is a lack of research about the promoting effect of vitamin C on the proliferation of adipose-derived stem cells under high glucose conditions and the related molecular mechanisms.OBJECTIVE:To explore the promoting effect of vitamin C on the proliferation adipose-derived stem cells treated by the high glucose and the related molecular mechanisms.METHODS:Passage 3 human adipose-derived stem cells were cultured under high glucose conditions and then treated with different concentrations of vitamin C (0, 100, 150, 200, 250, 300 μmol/L). Cells cultured under low glucose conditions acted as controls. The expression levels of p-ERK and p-AKT proteins were detected by western blot. MTT method was used to choose the optimal concentration and time of vitamin C for all the subsequent tests. Human adipose-derived stem cells cultured under high glucose conditions were divided into four groups, and cells in blank control group had no treatment. Cells in the other three groups were treated with the optimal concentration of vitamin C (vitamin C group), LY294002+the optimal concentration of vitamin C (LY294002 group), or U0126+the optimal concentration of vitamin C (U0126 group) for 48 hours.EdU staining assay was used to detect the cell proliferation of human adipose-derived stem cells.RESULTS AND CONCLUSION:(1) Cell counting kit detection:We found that high glucose reduced the proliferation of human adipose-derived stem cells, and vitamin C promoted the proliferation of these cells. The best concentration of vitamin C was 200 μmol/L and the optimal effect time was 48 hours. (2) Western blot detection:Compared with the 0 μmol/L vitamin C group, the level of p-ERK in the 200 μmol/L vitamin C group was upregulated significantly (P < 0.01),while no significant expression change in p-AKT protein was found in control, 0 and 200 μmol/L vitamin C groups.(3) EdU test:the number of EdU positive cells was significantly higher in the vitamin C, LY294002, and control groups compared with the blank control group (P < 0.01). Moreover, compared with the vitamin C group, the EdU positive cells in the U0126 group were decreased significantly in number (P < 0.01). In conclusion, the ERK/MAPK signaling pathway is involved in the promotion effect of vitamin C on the proliferation of human adipose-derived stem cells under high glucose conditions.