摘要
目的研究不同频率电刺激对L 929细胞(小鼠成纤维细胞)细胞周期和增殖情况的影响,进而探究不同频率的盆底电刺激在治疗轻中度女性压力性尿失禁(stress urinary incontinence, SUI)中可能的作用机制,为临床上采用物理电刺激方法治疗女性SUI提供相应的理论依据。方法采用本研究团队设计和制作的改良变频细胞电刺激装置对L 929细胞分别进行4组不同频率(25 Hz、50 Hz、75 Hz、100 Hz)的直流电刺激,电刺激时长为2 h,对照组不进行电刺激操作。电刺激干预结束后继续培养12 h,采用碘化丙啶染色和流式细胞术对细胞进行周期检测,并计算增殖指数。此外,在电刺激结束后采用EdU增殖试剂盒检测电刺激后细胞的增殖情况,荧光细胞计数后计算细胞的增殖率。结果与对照组相比,频率为25 Hz的电刺激使细胞的增殖指数增加,G0/G1期细胞数比例明显降低,S期和G2/M细胞数比例均较对照组明显增加,差异均有统计学意义(P<0.05)。采用频率为50 Hz电刺激时细胞的G0/G1期在总细胞数中的比例明显降低(P<0.05),且S期的细胞比例数较对照组明显增加(P<0.05);采用频率为75 Hz和100 Hz电刺激的细胞中,G0/G1期细胞数比例明显降低(P<0.05),且S期细胞比例数较对照组明显降低(P<0.05)。EdU增殖试验发现采用25 Hz和50 Hz的较低频率组电刺激比对照组细胞的增殖率明显增高(P<0.05),采用较高频率75 Hz和100 Hz的电刺激细胞的增殖率明显降低(P<0.05)。结论较低频率(25 Hz)的直流电刺激能直接促进成纤维细胞从相对静止的G0/G1期进入DNA合成期,加速细胞周期进程,促进细胞增殖。较低频率电刺激可能通过促进成纤维细胞增殖恢复女性盆底支持系统中细胞外基质的代谢和重构过程,帮助女性盆底功能的恢复。
Objective To investigate the effect of electrical stimulation with different frequencies on the cell cycle and proliferation of L 929 cells(mouse fibroblasts);to explore the possible mechanism of pelvic floor electrical stimulation with different frequencies in the treatment of stress urinary incontinence(SUI) for mild and moderate female patients;to provide the corresponding theoretical basis for clinical treatment of electrical stimulation in SUI. Methods L 929 cells were subjected to direct current stimulation at 4 different frequencies(25 Hz, 50 Hz, 75 Hz, 100 Hz) for 2 h with the modified variable frequency cell electrical stimulation device. No electrical stimulation was performed in the control group. After the electrical stimulation intervention, all cells were cultured for another 12 hours. The cell cycle was detected by propidium iodide staining and flow cytometry, and the proliferation index was calculated. In addition, EdU proliferation kit was used to detect the cell proliferation after electrical stimulation, and the cell proliferation rate was calculated after count. Results Compared with the control group, the proliferation index of L 929 cells was significantly increased in the 25 Hz electric stimulation group(P<0.01);the proportion of L 929 cells in the G0/G1 phase was significantly decreased(P<0.05);the proportion of L 929 cells in the S phase and G2/M phase was significantly increased(P<0.01). The proportion of cells in the G0/G1 phase was significantly decreased in the 50 Hz group(P<0.05), and the proportion of cells in the S phase was significantly increased compared with the control group(P<0.01). The proportion of cells in the G0/G1 phase was significantly reduced in the 75 Hz and 100 Hz electric stimulation groups(P<0.05);the proportion of cells in the S phase was significantly lower than that in the control group(P<0.05). The EdU proliferation test found that the proliferation rate of the cells in the lower frequency groups(25 Hz and 50 Hz) was significantly higher than that in the control group(P<0.05), and the proliferation rate of the cells in the higher frequency groups(75 Hz and 100 Hz) was significantly lower(P<0.05). Conclusion Lower frequency(25 Hz) direct current stimulation can directly promote fibroblasts from the relatively static G0/G1 phase to the DNA synthesis phase, accelerate the cell cycle process, and promote cell proliferation. Lower frequency electrical stimulation may help female pelvic floor function recovery by promoting fibroblast proliferation to restore the metabolism and remodeling of extracellular matrix in female pelvic floor support system.
作者
陈乾
洪莉
李洋
李素廷
CHEN Qian;HONG Li;LI Yang;LI Su-ting(Department of Obstetrics and Gynecology,Renmin Hospital of Wuhan Universily,Wuhan Hubei 430060,P.R.China)
出处
《中国计划生育和妇产科》
2019年第12期83-86,89,I0002,共6页
Chinese Journal of Family Planning & Gynecotokology
基金
国家自然科学基金资助项目(项目编号:81771562)
关键词
女性压力性尿失禁
电刺激治疗频率
成纤维细胞
细胞周期
细胞增殖
female stress urinary incontinence
electric stimulation frequency
fibroblast
cell cycle
cell proliferation
作者简介
通讯作者:陈乾,武汉大学人民医院妇产科硕士研究生在读,主要研究方向为女性盆底功能障碍性疾病和妇科肿瘤,E-mail:287375787@qq.com。
