Chen Chuan, Zhang Xiaogang, Dai Yanmei, Zhang Xuehui
Department of Cardiology, First Affiliated Hospital, Chongqing Medical University, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Apr;29(4):337-40.
To observe the inhibition of pulsed electric stimulation on the proliferation of H9c2 cells and the induction on the differentiation of the cells.
We applied a set of pulse electrical stimulations of different frequencies (0, 1, 5, 10, 50, 100 Hz), different voltage (0, 5, 10, 20, 40, 50 V) and different time (0, 1, 2, 4, 6, 8 h/d) on H9c2 cells in rats. 5-azacytidine was used as an intervention factor control. We detected the proliferation rate of H9c2 cells through MTT assay, the expressions of cardiac troponin T (cTNT) and Oct4 through immunofluorescence cytochemical staining, and the expressions of cTNT and α-MHC mRNA using RT-PCR method.
Pulsed electric stimulation inhibited the proliferation of H9c2 cells (P<0.05). The pulsed electric stimulation of 1 Hz, 4 h/d and 10 V was proved optimized in inhibiting the growth of H9c2 cells to the greatest extent but not inducing the death of cells. Two weeks after the optimized pulsed elctric stimulation on H9c2 cells, the expression of Oct4 was reduced (P<0.05), while the expressions of cTNT and α-MHC were still strong.
Pulsed electric stimulation can not only inhibit the reproduction of myoblast cell line H9c2 in heart tissue of rat embryo, but also induce the differentiation of H9c2 cells. During it, the expressions of cTNT and α-MHC are enhanced but Oct4 is weakened.
观察脉冲电刺激对H9c2细胞增殖的抑制作用及对细胞分化的诱导作用。
对大鼠H9c2细胞施加一组不同频率（0、1、5、10、50、100 Hz）、不同电压（0、5、10、20、40、50 V）和不同时间（0、1、2、4、6、8 h/d）的脉冲电刺激。以5-氮杂胞苷作为干预因素对照。通过MTT法检测H9c2细胞的增殖率，通过免疫荧光细胞化学染色检测心肌肌钙蛋白T（cTNT）和Oct4的表达，采用RT-PCR法检测cTNT和α-MHC mRNA的表达。
脉冲电刺激抑制了H9c2细胞的增殖（P<0.05）。证明1 Hz、4 h/d和10 V的脉冲电刺激在最大程度抑制H9c2细胞生长但不诱导细胞死亡方面效果最佳。对H9c2细胞进行优化的脉冲电刺激两周后，Oct4的表达降低（P<0.05），而cTNT和α-MHC的表达仍然很强。
脉冲电刺激不仅可以抑制大鼠胚胎心脏组织中成肌细胞系H9c2的增殖，还可以诱导H9c2细胞分化。在此过程中，cTNT和α-MHC的表达增强而Oct4减弱。
