脉冲电场对AC16人心室肌细胞的影响  

Effect of electricpulse field on AC16 human ventricular myocytes

作　　者：袁佳栎 李瑛泽 莫斌峰[1] 饶俊峰 王群山[1] YUAN Jia-li;LI Ying-ze;MO Bin-feng;RAO Jun-feng;WANG Qun-shan(Department of Cardiology,Xinhua Hospital Affiliated To Shanghai Jiaotong University School of Medicine,Shanghai 200092,China;School of Mechanical Engineering,University of Shanghai for Science and Technology)

机构地区：[1]上海交通大学医学院附属新华医院心内科,上海200092 [2]上海理工大学机械工程学院

出　　处：《中国心脏起搏与心电生理杂志》2021年第5期453-457,共5页Chinese Journal of Cardiac Pacing and Electrophysiology

基　　金：国家自然科学基金重大研究计划培育项目(91949128);面上项目(81873485);上海理工大学-上海交大医学院医工交叉重点支持项目(2021005)。

摘　　要：目的研究脉冲电场对AC16人心室肌细胞的影响。方法利用人心室肌AC16细胞系,使用ECM830电穿孔仪,设置脉宽1 ms、频率2 Hz/5 Hz、电场强度100~2 000 V/cm梯度脉冲电场对细胞进行电穿孔。再通过台盼蓝染色、CCK-8观察细胞活力,得到不同参数脉冲电场对AC16细胞活力的影响。并通过AnnexinV&PI法染色利用流式细胞术观察刺激后凋亡/坏死情况。结果在脉宽1 ms、2 Hz的基础脉冲参数作用后,观察到AC16细胞暴露在高场强的脉冲电场中细胞肿胀、破裂,发生不可逆性死亡。电压阈值大于1 000 V/cm时细胞死亡率明显增加(对照组:2%,1 000 V/cm组:36.8%),细胞活力明显降低(对照组的29.9%~46.4%)。且一定程度内增加脉冲次数与增加频率不改变引起死亡的电压阈值。Annexin V-FITC与PI共染色通过流式细胞术发现1 000及以上的电场强度显著增加了AnnexinV的荧光强度,证实1 ms的脉冲电场通过凋亡途径导致细胞死亡。结论 AC16细胞对脉冲电场诱导的凋亡具有稳定而明确的阈值,细胞死亡与脉冲次数和频率无关。Objective To study the effect of electricpulse field on the level of AC16 human ventricular myocytes. Methods We use human ventricular cardiomyocyte cell line AC16 and the ECM-830 electroporator was used to perform electroporation with pulse field under different parameters. Electroporation of cells was performed by setting pulse width of 1 ms, frequency of 2 Hz/5 Hz, and electric field intensity of 100-2 000 V/cm.Then, the activity of AC16 cells was observed by trypan blue staining and CCK-8, and the effects of different parameters of pulsed electric field on the activity of AC16 cells were obtained. The cell apoptosis or necrosis were measured by Annexin V & PI assay. Results After the basic pulse parameters with pulse width of 1 ms and 2 Hz, the AC16 cells were observed to swell and rupture in the electric pulse field with high field intensity, and irreversible death occurred. When the voltage threshold was greater than 1 000 V/cm, cell mortality was significantly increased(control group: 2%, 1 000 V/cm group: 36.8%) and cell viability was significantly decreased(control group: 29.9%-46.4%). The threshold of voltage causing death was not changed by increasing pulse frequency and frequency. Annexin V-FITC co-staining with PI showed that the fluorescence intensity of Annexin V was significantly increased by the electric field intensity of 1 000 and above by flow cytometry, which confirmed that the pulsed-electric field of 1 ms induced cell death through the apoptotic pathway. Conclusion The AC16 cells presented a stable and precise pulse field threshold that could lead to cell death.[Chinese Journal of Cardiac Pacing and Electrophysiology,2021,35(5):453-457]

关 键 词：心血管病学 脉冲电场 电穿孔 心脏消融 细胞凋亡 细胞坏死 

分 类 号：R454.1[医药卫生—治疗学]