Figure 4.

Mtp declines TBHP‐induced apoptosis and ROS production in BM‐EPCs. (A–C) ROS production detected by H₂ DCFDA fluorescence in BM‐EPCs. Cells were treated with 1, 10 and 100 μM Mtp for 48 hrs, labelled with H₂ DCFDA (30 μM) and then incubated with 25, 50 and 100 μM TBHP for 3 hrs prior to fluorescence microscopic analysis (scale bar: 200 μm). Mtp significantly attenuated TBHP‐induced ROS production; (D) live/dead staining results of cells treated by Mtp and/or TBHP. Cells were treated with 1, 10 and 100 μM Mtp for 48 hrs and TBHP for 3 hrs, followed by calcein‐AM/PI double staining. Cell survival was significantly up‐regulated by the preconditioning of Mtp even with TBHP treatment (scale bar: 200; 20 μm); (E) cell viability results of BM‐EPCs treated with Mtp and TBHP. Cell Counting Kit‐8 (CCK‐8) assay of BM‐EPCs pre‐treated with 1, 10 and 100 μM Mtp for 48 hrs followed by TBHP stimulation was performed, and cell viability was evidently increased by the Mtp pre‐treatment; (F, G) TUNEL assay was performed in BM‐EPCs as pre‐treated with 100 μM Mtp followed by TBHP stimulation, and Mtp ameliorates apoptosis of BM‐EPCs induced by TBHP (scale bar: 200 μm); (H, I) Western blot analysis results of levels of cleaved‐caspase 3 in BM‐EPCs with different doses of TBHP treatment. BM‐EPCs were treated with 25, 50 and 100 μM TBHP. The protein expression of cleaved‐caspase 3 was significantly increased after TBHP treatment; (J, K) Western blot analysis results of expression of cleaved‐caspase 3 after Mtp pre‐treatment. Cells pre‐treated with 100 μM Mtp followed by TBHP stimulation. Mtp reduced cleaved‐caspase 3 protein expression of BM‐EPCs induced by TBHP. The densitometric analysis of all Western blot bands was normalized to the total proteins or GAPDH. n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001 versus the indicated group.