中文摘要：

观察纳秒级陡脉冲电场诱导人浆液囊腺性卵巢癌SKOV3癌细胞凋亡效应,及其对细胞内钙离子浓度即[Ca2＋]i的影响并探讨其作用机理。采用场强为10kV/cm、脉宽为100ns、重复频率为1Hz的纳秒级陡脉冲电场作用于SKOV3癌细胞悬液5min。4h后采用Annexin V-FITC/PI双标记并结合流式细胞仪分析SKOV3癌细胞的凋亡率,采用扫描和透射电子显微镜观察癌细胞凋亡的超微结构变化;以Fluo-3/AM为细胞内钙离子的荧光指示剂,用激光扫描共聚焦显微镜检测纳秒级陡脉冲电场对SKOV3癌细胞[Ca2＋]i的影响,及[Ca2＋]i变化时的Ca2＋来源。结果显示,流式细胞术检测到癌细胞的早期凋亡率为（22.21±2.71）%,明显高于对照组的（3.04±0.44）%（P〈0.01）,扫描和透射电子显微镜均观察到典型的凋亡细胞形态,证实纳秒级陡脉冲电场能够有效诱导SKOV3癌细胞发生凋亡;同时纳秒级陡脉冲电场可明显升高细胞[Ca2＋]i（P〈0.01）,而与细胞外钙离子浓度无关（P〉0.05）。由于纳秒级陡脉冲电场的等值频率很高,容易穿透细胞膜,其诱导凋亡的机制之一可能是通过激活细胞内钙库（内质网、线粒体）,引起细胞内钙离子升高,从而介导凋亡信号通路。

英文摘要：

This paper investigated the apoptosis and changes of the intracellular calcium concentration（[Ca~（2＋）]i） induced by nanosecond pulsed electric field on human ovarian carcinoma cell Line SKOV3 in vitro.SKOV3 cells were exposed to nanosecond pulsed electric field（10kV/cm,pulse width 100ns,1Hz in 5minutes）.At 4h after treatment,the apoptosis of SKOV3 cells was detected by flow cytometry,SEM and TEM.The change of [Ca~（2＋）]i in SKOV3 cells and the source of Ca~（2＋） during [Ca~（2＋）]i change were observed by laser scanning confocal microscope（LSCM） using Fluo-3/AM as the calcium fluorenscent indicator.The results were shown that early apoptotic rate of experimental group was（22.21±2.71）%,significantly higher than that of the control group（3.04±0.44）%（P〈0.01）.The typical morphological features of apoptotic cells were observed by SEM and TEM.It was proved that nanosecond pulsed electric field can mainly induced early apoptosis.The [Ca~（2＋）]i was markedly increased by treatment with nanosecond pulsed electric field（P〈0.01）,while the increase showed no relationship to the extracellular calcium concentration（P〉0.05）.The equivalent frequency of nanosecond pulsed electric field is much larger,which can easily penetrate into the cell membrane,one of the mechanisms of the apoptosis may be that nanosecond pulsed electric field can activate intracellular calcium and result in increase of [Ca~（2＋）]i soas to induce apoptosis of SKOV3 cells.