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目的:旨在通过检测电刺激干预骨骼肌卫星细胞(C2C12)后细胞活性与肌球蛋白重链的表达,阐明骨骼肌卫星细胞增殖与分化的适宜条件,为临床治疗体育运动造成的骨骼肌损伤提供新的思路。方法:CPace EP电刺激仪按不同电压、频率、刺激时间、脉冲时间条件施加刺激,MTT法检测细胞活性,Western blot法检测MYH蛋白含量。结果:1)与对照组相比,电压≤10 V、刺激时间≤150 min、频率≤5.0 Hz、2 ms≤脉冲时间≤4 ms时,各实验组吸光度无统计学差异。2)对照组MYH蛋白含量均较低,5V、10V、15V组MYH蛋白含量均上升,其中5V组增加最显著;45 min、90 min、180 min组细胞内MYH蛋白含量均增加明显,其中180 min组MYH含量最高;1 Hz、7.5 Hz、15 Hz组MYH蛋白含量均明显增加,其中1 Hz组增加最显著;,1 ms组和2 ms组C2C12细胞内MYH蛋白含量有明显的上升,2 ms组增加最显著,16 ms组MYH无明显变化。结论:电刺激C2C12细胞时,其条件以电压≤10 V、刺激时间≤150 min、频率≤5.0 Hz、2 ms≤脉冲时间≤4 ms为宜,细胞活性不受明显影响。适宜的电刺激能够促进C2C12细胞的分化,其适宜条件分别为电压5V、刺激时间180 min、频率1Hz、脉冲时间2 ms。
OBJECTIVE: To detect the cell viability and myosin heavy chain expression after skeletal muscle satellite cells (C2C12) stimulation by electrical stimulation and to elucidate the appropriate conditions for the proliferation and differentiation of skeletal muscle satellite cells for the treatment of skeletal muscle injury caused by physical exercise Provide new ideas. Methods: CPace EP electrostimulator was used to stimulate cells under different voltage, frequency, stimulation time and pulse time. Cell viability was measured by MTT assay. MYH protein content was detected by Western blot. Results: 1) Compared with the control group, the absorbance of each experimental group had no statistical difference when the voltage ≤10 V, the stimulation time ≤150 min, the frequency ≤5.0 Hz, 2 ms≤ pulse time ≤4 ms. 2) The content of MYH in control group was lower, and the content of MYH in 5 V, 10 V, 15 V group increased, among which, the increase in 5 V group was the most significant. The content of MYH protein in 45 min, 90 min and 180 min groups increased significantly The content of MYH in 1 min, 7.5 Hz and 15 Hz groups were significantly increased, among which 1 Hz increased most significantly; MYH protein content in C2C12 cells in 1 ms and 2 ms group increased obviously, 2 ms group increased the most significant, 16 h MYH group no significant change. CONCLUSION: When C2C12 cells were electrically stimulated, the conditions were as follows: voltage ≤10 V, stimulation time ≤150 min, frequency ≤5.0 Hz, 2 ms ≤ pulse time ≤4 ms, and cell activity was not significantly affected. Appropriate electrical stimulation can promote the differentiation of C2C12 cells. The suitable conditions are voltage 5V, stimulation time 180 min, frequency 1 Hz, pulse time 2 ms.