Adrenomedullin Gene Transfected Bone Marrow Mesenchymal Stem Reduce Apoptosis Under Hypoxia-cultured Conditions

[Background] Following transplantation, bone marrow mesenchymal stem cells (BMSCs) were in a microenvironment of hypoxia and ischemia in the ischemia/infarct myocardium. Ad-ADM transfection enhanced the effects of MSCs transplantation.[Objective] To investigate the proliferation, apoptosis and expression of vascular endothelial growth factor (VEGF) in BMSCs after adrenomedullin(ADM) transfection under hypoxia and serum-free culture conditions.[Methods] Rat BMSCs were conventionally harvested by differential adhesion method. The infection efficiency of adenovirus vector to MSCs was tested by x-gal stain. MSCs were divided into control group, Ad-vector transfected MSCs group and ADM gene transfected MSCs group. MSCs cultured under hypoxia and serum-free conditions for 0,3,6,9,12,16,20,24h respectively.Cell proliferation was detected by cell counting kit-8 (CCK-8).ADM and VEGF in culture medium ware detected by enzyme-linked immunosorbent (ELISA).Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick labeling (Tunel).[Results] Infection rate of MSCs increased with MOI in a dose-dependant manner. When MOI was 150, the infection rate was 95.4%. After MSCs cultured in hypoxia and serum-free medium, cell growth inhibition and apoptosis increased in a time dependent manner, but the apoptosis rate in ADM gene transfected MSCs group were significantly lower than other 2 groups after cultured 0,3,6,9,12,16 h(all P<0.05). At the same time, MSCs secrete ADM and VEGF increased and ADM gene transfection MSCs group were significantly higher than other 2 groups.[Conclusion] Under hypoxia and serum-free cultured conditions, ADM gene transfected MSCs enhance the ability of anti-apoptotic, which may be related to the increased expression of ADM and VEGF of MSCs.