Figure 7.

AdC decreases COX2 methylation and attenuates down HMSCs senescence. (A-C) COX2 methylation was decreased after HMSCs were treated with 1 µM AdC. ^*P<0.05 as compared with the G10, G15 or SIPS group. (D and E) COX2 protein quantities were detected by western blot analysis and the expression levels were calculated. Statistical analysis results showing that the COX2 expression level increased after HMSCs were treated by AdC in both RS and SIPS cells. ^**P<0.01 as compared with the G15 or SIPS group. (F-H) WST-1 assay showing that AdC treatment led to the more rapid growth of G5, G10 and G15 HMSCs than those untreated at the same generation. The analysis time points were 24 and 48 h. ^*P<0.05 and ^**P<0.01 as compared with the CT groups. (I-L) SA-β-Gal staining results showing that AdC attenuates HMSC senescence. (I) HMSCs in the RS model; (J) HMSCs in the SIPS model; (K and L) statistical analysis results of HMSCs stained positive for SA-β-Gal. ^**P<0.01 as compared with the RS (G15) and SIPS (14 days) groups shown in Fig. 1C and F. HMSCs, heart mesenchymal stem cells; RS, replicative senescence; SIPS, stress-induced premature senescence; AdC, 5-aza-2′-deoxycytidine.