Abstract
Objective To investigate the effect of AKT gene modified amniotic fluid- derived mesenchymal stem cells (AF-MSC) on oxidative stress after transplantation in rabbit models with myocardial ischemia-reperfusion injury (IRI). Methods New Zealand rabbits were randomly divided into three groups, Group A: low glucose-Dulbecco's modified Eagle's medium (L-DMEM) group, Group B: AF-MSC group, and Group C: AKT-AFMSC group. The myocardial IRI rabbit models were established. Groups L-DMEM, AF - MSC, and AKT - AFMSC received direct injection in myocardial infarction area and surrounding membrane before reperfusion, respectively. The arterial blood and injured myocardium were included at 2 hours and 21 days after the operation, respectively. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were measured by Xanthine oxidase and Thiobarbituric acid (TBA) assay, respectively. ELISA was used to measure 3-nitrotyrosine (3-NT) and nitric oxide (NO) content. Results At 2 hours after reperfusion, the comparison between AKT- AFMSC transplantation group versus L- DMEM control group and AF-MSC transplantation group showed significantly decreased MDA content (2.01±0.53 vs. 4.98±0.90 and 4.46±0.84, P<0.01) , NO content (12.38±1.54 vs. 19.15±3.40 and 18.47±7.22, P<0.01) , 3- NT content (204.87±19.16 vs. 237.32±23.05 and 241.36±13.31, P<0.01) , and increased SOD activity (185.72±18.42 vs. 133.98±20.06 and 129.89±15.68, P<0.01) , and the differences were statistically significant. At 21 days after the operation, the comparison between AF-MSC transplantation group and AKT- AFMSC transplantation group versus L-DMEM control group showed significantly decreased myocardial MDA content (0.21±0.03 and 0.09±0.02 vs. 0.26±0.04, P<0.05, P<0.01, respectively) , NO content (6.04±0.31 and 5.43 0.37 vs. 6.90±0.39, P<0.05, P<0.01, respectively) , 3-NT content (180.26±7.65 and 158.68±8.81 vs. 195.95±13.29, P<0.05, P<0.01, respectively) , and increased SOD activity (14.55±0.46 and 17.95±0.86 vs. 13.18±0.75, P<0.05, P<0.01, respectively) , and the differences were statistically significant. Conclusion In myocardial IRI, the transplanted AKT gene modified AF-MSC can significantly reduce MDA, NO, 3-NT levels and increase SOD activity, which has stronger anti-oxidative stress than AF- MSC. Key words: Amniotic fluid; Mesenchymal stem cells; Ischemic reperfusion injury
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