摘要目的 探讨AKT基因修饰的羊水间充质干细胞(AF-MSC)移植于兔缺血再灌注损伤心肌模型后对其氧化应激状态的影响.方法 将新西兰兔随机分成3组,A组:L-DMEM组、B组:AF-MSC(羊水间充质干细胞)组、C组:AKT-AFMSC(AKT-羊水间充质干细胞)组,建立兔心肌缺血再灌注损伤模型.再灌注开通前在梗死区及周边心肌外膜按分组类别直接注射:L-DMEM、AF-MSC和AKT-AFMSC.术后2h收集动脉血、术后21 d收集损伤心肌,采用黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活力,硫代巴比妥酸(TBA)法检测丙二醛(MDA)含量.ELISA方法检测3-硝基酪氨酸(3-NT)、一氧化氮(N0)含量.结果 在再灌注后2h,AKT-AFMSC移植组与L-DMEM对照组和AF-MSC移植组比较:血清MDA含量(2.01±0.53比4.98±0.90和4.46±0.84)均显著降低(P＜0.01)、NO含量(12.38±1.54比19.15±3.40和18.47±7.22)均显著降低(P＜0.01)、3-NT含量(204.87±19.16比237.32±23.05和241.36±13.31)均显著降低(P＜0.01),SOD(185.72± 18.42比133.98±20.06和129.89±15.68)活力显著升高(P＜0.01),差异有统计学意义.术后21d,AF-MSC移植组和AKT-AFMSC移植组:较L-DMEM对照组心肌MDA含量降低(0.21±0.03和0.09±0.02比0.26±0.04)分别为P＜0.05,P＜0.01、NO含量降低(6.04±0.31和5.43±0.37比6.90±0.39)分别为P＜0.05,P＜0.01、3-NT含量降低(180.26±7.65和158.68±8.81比195.95±13.29)分别为P＜0.05,P＜0.01、而SOD活力升高(14.55±0.46和17.95±0.86比13.18±0.75)分别为P＜0.05,P＜0.01,差异有统计学意义,并且AKT-AFMSC移植组较AF-MSC移植组心肌MDA、NO、3-NT含量降低更明显(P＜0.01),SOD活力升高更明显(P＜0.01),两组相比较差异有统计学意义.结论 在缺血再灌注损伤心肌中,移植AKT基因修饰的羊水间充质干细胞能显著降低MDA、NO、3-NT水平,增加SOD活力,且较羊水间充质干细胞具有更强的抗氧化应激作用.

abstractsObjective 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 AKTAFMSC 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.