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目的观察不同强度及作用时长的电刺激对小鼠成纤维细胞L 929细胞活性、衰老及凋亡的影响。方法实验于2016年7~10月在武汉大学人民医院中心实验室进行。根据电刺激时长的不同,将小鼠成纤维细胞L 929细胞分为0 h组、2 h组、4 h组、6 h组,其中0 h组为对照组;根据电刺激强度的差异再将每组细胞分为50 mV/mm、100 mV/mm、200 mV/mm组,共计12组。采用CCK-8试剂盒、细胞衰老β-半乳糖苷酶染色试剂盒、线粒体膜电位检测试剂盒(JC-1)检测其活性、衰老及凋亡,每组实验重复4次。结果与对照组相比,50 mV/mm 4 h与6 h组、100 mV/mm 2 h组细胞活性增高(P<0.05),而50 mV/mm 2 h组、100 mV/mm 6 h组、200 mV/mm 2 h与6 h组细胞活性下降(P<0.05),100 mV/mm 4 h组、200 mV/mm 4 h组细胞活性变化不明显;100 mV/mm 2 h组细胞衰老率降低(P<0.05),而50 mV/mm 2 h、4 h、6 h组、100 mV/mm 6 h组、200 mV/mm 4 h与6 h组细胞衰老率升高(P<0.05),100 mV/mm 4 h组、200 mV/mm 2 h组细胞衰老率变化不明显;50 mV/mm 2 h组与100 mV/mm 2 h组凋亡细胞比例下降,其余组凋亡细胞比例均升高。结论小鼠成纤维细胞L 929细胞在不同的电场强度及作用时长下活性、衰老、凋亡状态各不相同,因此在研究电刺激对细胞的影响时,应充分考虑电刺激的强度及作用时间。其中,100 mV/mm 2 h的直流电刺激可以提高小鼠成纤维细胞L 929细胞的活性,抑制其衰老及凋亡。
Objective To observe the effects of different intensities and duration of electrical stimulation on the activity, aging and apoptosis of mouse fibroblast L 929 cells. Methods The experiment was performed in the Central Laboratory of Wuhan University People’s Hospital from July to October 2016. According to the different duration of electrical stimulation, mouse fibroblasts L 929 cells were divided into 0h group, 2h group, 4h group, 6h group, 0h group as the control group; according to the difference of electrical stimulation intensity then Each group of cells was divided into 50 mV / mm, 100 mV / mm, 200 mV / mm group, a total of 12 groups. CCK-8 kit, cell senescence β-galactosidase staining kit and mitochondrial membrane potential detection kit (JC-1) were used to detect the activity, aging and apoptosis. Each experiment was repeated 4 times. Results Compared with the control group, the cell viability increased at 50 mV / mm 4 h and 6 h and 100 mV / mm 2 h (P <0.05), while at 50 mV / mm 2 h and 100 mV / mm 6 h (P <0.05). The cell viability in 200 mV / mm 4 h group and 200 mV / mm 4 h group did not change significantly. The cell senescence in 100 mV / mm 2 h group (P <0.05). The cell aging rate in 50 mV / mm 2 h, 4 h, 6 h, 100 mV / mm 6 h and 200 mV / mm 4 h and 6 h groups increased ), 100 mV / mm 4 h group and 200 mV / mm 2 h group, the percentage of apoptotic cells decreased in 50 mV / mm 2 h group and 100 mV / mm 2 h group. The percentage of apoptotic cells in the other groups Proportion increased. CONCLUSION: The activity, aging and apoptosis status of L929 cells in different time and in different electric fields are different. Therefore, in studying the effect of electrical stimulation on cells, the intensity and duration of electrical stimulation should be fully considered . Among them, 100 mV / mm 2 h DC stimulation can increase the activity of mouse fibroblast L 929 cells, inhibit its aging and apoptosis.