论文部分内容阅读
骨髓间充质干细胞(BMSCs)由于具有向心肌方向分化的潜能,目前已成为一种新的能够修复心脏组织损伤的重要治疗选择。在本实验中我们探讨了体外不同的电刺激时间对BMSCs向心肌方向分化的影响。实验共分为三组:1电刺激组,完全培养基培养的大鼠骨髓间充质干细胞(rBMSCs)受到电压为2V,频率为2 Hz,脉宽为5ms的电刺激,作用时间为30min/d、2h/d、4h/d、6h/d,连续作用10d;25-氮胞苷(5-Aza)组,5-Aza(10μmol/L)诱导rBMSCs 24h,然后采用完全培养基培养10d;3空白对照组,完全培养基静态培养rBMSCs 10d。通过实时荧光定量PCR方法检测各组rBMSCs的肌细胞特异性增强因子-2C(MEF-2C)mRNA和缝隙连接蛋白43(Cx43)mRNA的表达;通过Western blot方法检测各组rBMSCs的MEF-2C蛋白和Cx43蛋白的表达。实验结果显示:不同持续时间的电刺激组及5-Aza组的MEF-2CmRNA、Cx43mRNA表达量及MEF-2C蛋白、Cx43蛋白表达量均明显高于空白对照组(P<0.05)。本实验结果提示,体外电刺激可以诱导rBMSCs向心肌方向分化,其中2h/d作用效果最佳,但其机制尚未明确。
Bone marrow mesenchymal stem cells (BMSCs) have become a new important treatment option for repairing heart tissue damage due to their potential to differentiate into cardiac muscle. In this experiment, we explored the different in vitro electrical stimulation of the BMSCs to the direction of myocardial differentiation. The experiment was divided into three groups: 1, electrical stimulation group, rat bone marrow mesenchymal stem cells (rBMSCs) cultured in complete medium were subjected to electrical stimulation at a voltage of 2V, a frequency of 2 Hz and a pulse width of 5 ms for 30 min / The rBMSCs were induced by 25-azacytidine (5-Aza) and 5-Aza (10μmol / L) for 24h, then cultured in complete medium for 10 days. 3 blank control group, complete medium static culture rBMSCs 10d. The mRNA expression of MEF-2C and connexin 43 (Cx43) in rBMSCs of each group was detected by real-time fluorescence quantitative PCR. MEF-2C protein of rBMSCs was detected by Western blot And Cx43 protein expression. The results showed that the expressions of MEF-2C mRNA, Cx43 mRNA, MEF-2C protein and Cx43 protein in electrical stimulation group and 5-Aza group at different durations were significantly higher than those in blank control group (P <0.05). The experimental results suggest that in vitro electrical stimulation can induce rBMSCs to differentiate into the direction of myocardium, of which 2h / d has the best effect, but its mechanism is not yet clear.