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目的 探讨羊水来源间充质干细胞(AFMSC)诱导分化为心肌细胞的可行性.方法 抽取新西兰孕兔羊水分离培养AFMSC.选取八聚体结合转录因子4(Oct4)蛋白表达阳性的AFMSC分为实验组和对照组,实验组使用5-氮胞苷处理细胞24h,对照组加入等量磷酸盐缓冲液(PBS).处理后1、2、3、4周使用实时PCR检测心肌特异性转录因子GATA4 mRNA表达,免疫印迹法检测心肌肌钙蛋白T(cTnT)蛋白表达.结果 5-氮胞苷处理AFMSC后1、2、3周,实验组与对照组GATA4 mRNA和cTnT蛋白表达差异无统计学意义(均P＞0.05).与对照组比较,5-氮胞苷处理4周实验组GATA4 mRNA和cTnT蛋白表达均显著增加(GATA4:4.102±1.203比2.230± 1.010,cTnT:0.89±0.06比0.74±0.09,均P＜0.05).结论 AFMSC具有多能干细胞特性,经5-氮胞苷处理后可以向心肌细胞诱导分化.

Objective To investigate the feasibility of induced transformation of cardiomyocytes from amniotic fluid-derived mesenchymal stem cells (AFMSCs).Methods AFMSCs isolated from amniotic fluid of pregnant New Zealand rabbits were cultured,and those with positive expression of octamer-binding transcription factor 4 (Oct4) were divided into experiment group and control group.The experiment group was induce by 5-azacytidine for 24 h,and the control group was added with equal volume of phosphatebuffered solution (PBS).At 1,2,3 and 4 weeks after the treatment,the mRNA expression of cardiacspecific transcription factor GATA4 mRNA was assayed by real-time PCR,and the expression of cardiac troponin T (cTnT) was detected by Western blotting.Results At 1,2 and 3 weeks after treatment with 5-azacytidine,there were no differences in expression of GATA4 mRNA and cTnT between the experiment group and control group (allP＞0.05).However,at week 4,the expression of GATA4 mRNA (4.102±1.203 vs 2.230± 1.010) and cTnT (0.89±0.06 vs 0.74±0.09) in experiment group increased significantly compared with the control group (both P＜0.05).Conclusion AFMSC exhibits the characteristics of pluripotent stem cells,and may be differentiated to myocardial cells after induced with 5-azacytidine.