高强度皮秒脉冲电场抑制HUVEC迁移、增殖及小管形成的实验研究

Experimental Study on Inhibition Effect of HUVEC Migration, Proliferation and Tube Formation by Intense Picosecond Pulsed Electric Field

目前,国内外关于皮秒脉冲电场(picosecond pulsed electric field,ps PEF)治疗肿瘤的研究多集中于ps PEF对肿瘤的杀伤效应,对肿瘤所赖以生存的血管生成的研究鲜有提及。为此,采用脉冲宽度为800 ps,重复频率为3 Hz,脉冲个数为2 000,电场强度分别为20 MV/m、30 MV/m,40 MV/m的皮秒脉冲电场处理人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC),利用划痕实验、Transwell小室法、cell counting kit-8(CCK-8)法及体外小管形成实验,初步探究了ps PEF对血管生成的影响。实验结果表明:ps PEF可以有效降低HUVEC的迁移率和增殖率,且存在明显的剂量依赖关系,细胞迁移抑制率、细胞增殖抑制率均随ps PEF的增强而逐渐增大,其中细胞增殖抑制率在脉冲处理12 h后达峰值,细胞迁移和增殖抑制率最大可达73.68%,95.54%(40 MV/m);ps PEF可抑制HUVEC小管的形成,ps PEF场强越大,脉冲处理后小管形成抑制率越高,最高可达95.83%(40MV/m)。实验结果初步验证了高强度ps PEF对血管生成的抑制效果。

To preliminarily explore the impact of picosecond pulsed electric field on angiogenesis, psPEF with pulse width 800 ps, repetition frequency 3 Hz, pulse number 2 000, the field intensity 20 MV/m, 30 MV/m, 40 MV/m was delivered to human umbilical vein endothelial cells（HUVEC）. Scratch test, Transwell chamber method, cell counting kit-8 （CCK-8）, and tube formation experiment were used to preliminarily explore the impact of picosecond pulsed electric field on angiogenesis. The results show that picosecond pulsed electric field can effectively reduce cell migration rate and cell proliferation rate of human umbilical vein endothelial cells with significant dose-dependent relationship. With the in- creasing of picosecond pulsed electric field, the inhibition rates of cell migration and cell proliferation gradually increase. And the inhibition of cell proliferation reaches the maximum value 12 hour after picosecond pulsed electric field treatment. The inhibition rates of cell migration and cell proliferation can be up to 73.68% and 95.54% （40 MV/m）, respectively. Picosecond pulsed electric field can inhibit the tube formation rate of human umbilical vein endothelial cells. The stronger the picosecond pulsed electric field is, the larger the inhibition of tube formation rate after psPEF treatment is,even reaching its maximum extent of 95.83% （40 MV/m）. Research results can preliminarily verify the inhibitory effects of picosecond pulsed electric field on angiogenesis of HUVEC.