Research of Adaptive Protection of Preconditioning with Hydrogen Peroxide in Bone Marrow Mesenchymal Cells Against Apoptosis / 中山大学学报(医学科学版)

ABSTRACT

[Objective] To investigate the effects of preconditioning with low concentration of hydrogen peroxide (H2O2) on oxidative stress-induced BMSC apoptosis.[Methods] In vitro separation,purification,culture,and amplification of bone marrow mesenchymal stem cells were performed.BMSC were insulted with 0,50,100,200,300,400,500 μmol/L H2O2 and the effect of different consentration of H2O2 on BMSC was detected by Flow cytometry (FCM).And then cells were preconditioned with different consentraion of H2O2.(FCM) was used to determine the protective role of H2O2 preconditioning on BMSC apoptosis,BMSC chromatin distribution changes were observed by Hoechst33324;BMSC Caspase-3 and Bcl-2 gene changes were detected by RT-PCR.[Results] Analysis of BMSC apoptosis by flow cytometry showed that H2O2 induced BMSC apoptosis in a dose-dependent manner,and pretreatment of the cells with low concentration of H2O2 prevented subsequent stimulation with high H2O2.RT-PCR results showed that preconditioning with low concentration of H2O2 reduced the BMSC Caspase-3 gene expression but increased Bcl-2 gene expression.[Conclusion] Preconditioning with low concentration of H2O2 has an adaptive role in BMSC,and its mechanism may be related to inhibit abnormal gene expression of Caspase-3 and increase the gene expression of Bcl-2.