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目的探讨低频脉冲电磁场(PEMFs)对大鼠骨髓间充质干细胞(rBMSCs)α-辅肌动蛋白-2(ACTN2)、α-肌动蛋白(α-actin)和肌钙蛋白T-2(TNNT2)表达的影响。方法采用差速贴壁分离方法获取rBMSCs,选用第3代rBMSCs用于实验。实验分为4组:①PEMFs诱导组,给予50Hz、1mT低频PEMFs刺激,30min/d,分别刺激10d、15d和20d;②5-氮胞苷(5-Aza)诱导组,给予10μmol/L 5-Aza诱导1d,然后采用完全培养液培养10d、15d和20d;③PEMFs+5-Aza诱导组:采用10μmol/L 5-Aza诱导1d后,再给予PEMFs诱导组相同的处理;④对照组,采用完全培养液培养10d、15d和20d。通过倒置相差显微镜逐日连续观察细胞的生长状况和形态特征。采用实时荧光定量PCR方法检测ACTN2和TNNT2的mRNA表达,采用Western blot方法检测ACTN2、α-actin和TNNT2的蛋白表达。结果在各时间点,PEMFs诱导组、5-Aza诱导组和PEMFs+5-Aza诱导组的ACTN2、α-actin和TNNT2mRNA表达与蛋白表达明显高于对照组。结论 PEMFs可促进体外培养的rBMSCs向心肌样细胞分化。
Objective To investigate the effect of low frequency pulsed electromagnetic fields (PEMFs) on the expression of ACTN2, α-actin and Troponin T-2 (TNNT2) in rat bone marrow mesenchymal stem cells ) Expression. Methods The rBMSCs were obtained by differential adherent separation and the third generation rBMSCs were selected for experiment. The experiment was divided into 4 groups: ①PEMFs induced group, given 50Hz, 1mT low frequency PEMFs stimulation, 30min / d, respectively, stimulated 10d, 15d and 20d; ② 5-azacytidine (5-Aza) induction group, given 10μmol / L 5-Aza The cells were cultured in complete medium for 10 days, 15 days and 20 days, respectively; ③PEMFs + 5-Aza induction group: treated with 10μmol / L 5-Aza for 1 day, then treated with PEMFs induction group; ④The control group, The liquid culture 10d, 15d and 20d. The growth and morphological characteristics of the cells were observed continuously by inverted phase contrast microscope. The mRNA expressions of ACTN2 and TNNT2 were detected by real-time fluorescence quantitative PCR, and the protein expressions of ACTN2, α-actin and TNNT2 were detected by Western blot. Results At each time point, the expressions of ACTN2, α-actin and TNNT2 mRNA in PEMFs-induced group, 5-Aza-induced group and PEMFs + 5-Aza induced group were significantly higher than those in control group. Conclusion PEMFs can promote the differentiation of rBMSCs into cardiomyocyte in vitro.