| External factors: | Melatonin |
| Aging type: | Prevent |
| Aging characteristic: |
| Category: | Chemical compounds |
| Phenotype: | Aging |
| Experiment: | SA-β-gal activity assay//Flow cytometry//Immunofluorescence |
| Description: | After treatment with 100 μM H2O2 and incubation for 24 h,cells were stained with SA-βgal staining solution.We found that the number of SA-βgal-positive cells increased significantly compared to the untreated-control cells. Furthermore, the number of SA-βgal-positive cells treated with both 100 μM H2O2 and 1 μM melatonin was significantly lower than the H2O2-treated group .To confirm aging, a cell cycle assay and flow cytometry analysis were performed and revealed that treatment with H2O2 caused a change in the cell cycle phase distribution,representing cell cycle arrest. Cells treated with H2O2 showed a significantly higher percentage of cells in the subG1 phase and a lower proportion in the S phase compared to untreated-control cells. In addition, H2O2-melatonin-treated cells demonstrated significantly decreased subG1 phase, which represented the cell cycle arrest state, compared with H2O2-treated cells.Ki67 expression was used to detect alterations in proliferation.Melatonin increased Ki67 expression, whereas cells preincubated with H2O2 demonstrated decreased Ki67 expression. Treatment with H2O2 and melatonin resulted in an increase in the number of Ki67-positive cells compared to cells treated with H2O2 alone. |
| Regulatory pathway: | SIRT1 |
| R-EF-Pathway: | Upregulation |
| Official symbol(s): | SIRT1 |
| Pathway experiment: | Western blot |
| Pathway description: | These results showed that SIRT1 levels of cells treated with 100 μM H2O2 alone were significantly decreased compared to untreated-control cells. In cells co-treated with 1 μM melatonin, SIRT1 levels were rescued to 91.3 ± 3.1% (p < 0.01) of control levels compared to the H2O2-treated group and increased in melatonin-treated alone cells.In control cells, there was moderate SIRT1 immunoreactivity (A). Confocal analysis revealed the formation of SIRT1-positive aggregates in melatonin-treated cells (B). |
Annotation: