电刺激对体外培养C_2C_(12)细胞抗氧化能力的影响

Effect of Electric Stimulation on Anti-oxidation Capacity of C_2C_(12) Cell in Vitro

目的:通过电刺激体外培养的骨骼肌卫星细胞(C2C12),观察其对细胞活性和抗氧化能力的影响,探索适宜的体外电刺激条件及细胞抗氧化能力改变的机制。方法:C-Pace EP电刺激仪刺激体外培养的C2C12细胞,MTT法检测细胞活性,DCF法检测细胞内ROS含量,Western blot法检测细胞核内Nrf2蛋白含量。结果:1)0-40.0 V电压刺激细胞,各组细胞相对吸光度值分别为1.000、0.993、0.969、0.968、0.954、0.947、0.836、0.798;0-20.0 Hz频率刺激后,各组细胞相对吸光度值分别为1.000、1.022、1.023、1.055、1.055、1.154、1.140、1.104、1.102。2)与对照组比较,15 V电刺激细胞内相对DCF荧光强度显著增强。3)对照组细胞核内Nrf2蛋白含量均较低,电刺激后含量显著增加,5 V和15 Hz组分别是所检测不同电压和频率条件下促Nrf2入核作用最强组。结论:体外培养C2C12细胞进行电刺激干预时,要考虑电刺激的电压和频率,适宜的电刺激可通过促进Nrf2入核增强细胞的抗氧化能力。

Objective： This study observed the effect of electric stimulation on cell activity and anti-oxidation capac-ity of C2C12 cell in vitro, and explored the suitable condition of electric stimulation and the mechanism of change ofanti-oxidative capacity. Methods： C2 Cn cell was cultivated and stimulated by C-Pace EP. Cell activity of C2 C12 cellwas tested by MTT, the content of reactive oxygen species （ROS） of C2C12 cell was tested by DCF and the proteincontent of Nrf2 in nucleus of C2 C12 cell was tested by western blot. Results： 1 ） Compared with control group, therelative absorbance of each group stimulated by different voltage （0 -40. 0 V） is 1. 000, 0. 993, 0. 969, 0. 968,0. 954, 0. 947, 0. 836, and 0. 798 respectively. When stimulated by different frequency （0 -20. 0 Hz）, the rela-tive absorbance of each group is 1. 000, 1. 022, 1. 023, 1. 055, 1. 055, 1. 154, 1. 140, 1. 104, and 1. 102 re-spectively. 2） Compared with control group, relative DCF fluorescence intensity in other groups is improved signif-icantly after 15 V electric stimulation. 3 ） The protein level of Nrf2 in cell nucleus is low in control group, and it＇s increased/ffter electric stimulation. The groups stimulated by 5 V electric and 15 V electric are the most powerfulgroups to promote Nrf2 into cell nucleus. Conclusion： The voltage and frequency of electric stimulation should beconsidered when using electric to stimulate C2C12 cell. Suitable electric stimulation could enhance cell＇ s anti-oxi-dation capacity by promoting Nrf2 into cell nucleus.