Figure 3.

Melatonin prevented H₂O₂-induced premature senescence through the AMPK-SIRT1 signaling pathway. OVX BMMSCs were first exposed to H₂O₂ (100 μM) for 2 h and then treated with melatonin (MT) at 1 μM or 100 μM for an additional 72 h. (a) The mRNA expression levels of Sirt1, P16, P21, and P53 were quantified. (b, c) The protein levels of p-AMPK, AMPK, SIRT1, P16, P21, and P53 were determined using Western blot assays. (d) To inhibit the phosphorylation of AMPK, OVX BMMSCs were pretreated with 10 μM of compound C (CC) and then treated with 100 μM of melatonin. The mRNA expression levels of Sirt1 were quantified using qRT-PCR. (e, f) The protein levels of p-AMPK, AMPK, and SIRT1 were determined using Western blot assays. Values are presented as the mean ± S.E.M of four independent experiments (n = 4) in qRT-PCR experiments and three independent experiments (n = 3) in Western blot assays. Statistically significant differences are indicated by ^∗p < 0.05 or ^∗∗p < 0.01 between the indicated groups.