CAJ | 학술논문

为研究高强度ps脉冲电场（psPEF）诱导HeLa细胞的生物电效应，将脉冲电场参数组合（电场强度为250kV／cm，脉宽为800ps，脉冲个数为1000、3000、5000，频率为3Hz）作用于人宫颈癌HeLa细胞。利用噻唑蓝（methylthiazolyltetrazolium，MTT）比色法检测ps脉冲对细胞的生长抑制情况；钙离子指示剂（Fluo-3／AM）探针标记细胞，激光共聚焦显微镜检测细胞内钙离子体积分数的改变；蛋白质印迹法检测促凋亡蛋白Bax与凋亡抑制蛋白Bcl-2释放水平的改变。实验发现：细胞死亡率与施加脉冲个数正相关，且处理后12h抑制率最高；激光扫描共聚焦半定量分析显示，处理组细胞荧光强度明显高于对照组（检验水准P〈0．05）；蛋白质印迹法检测结果表明，处理组细胞内Bax表达量增加（P〈0．05），Bcl-2释放量略有降低。上述结果表明：高强度ps脉冲通过诱导细胞凋亡进而抑制了HeLa细胞的增殖。
위연구고강도ps맥충전장（psPEF）유도HeLa세포적생물전효응，장맥충전장삼수조합（전장강도위250kV／cm，맥관위800ps，맥충개수위1000、3000、5000，빈솔위3Hz）작용우인궁경암HeLa세포。이용새서람（methylthiazolyltetrazolium，MTT）비색법검측ps맥충대세포적생장억제정황；개리자지시제（Fluo-3／AM）탐침표기세포，격광공취초현미경검측세포내개리자체적분수적개변；단백질인적법검측촉조망단백Bax여조망억제단백Bcl-2석방수평적개변。실험발현：세포사망솔여시가맥충개수정상관，차처리후12h억제솔최고；격광소묘공취초반정량분석현시，처리조세포형광강도명현고우대조조（검험수준P〈0．05）；단백질인적법검측결과표명，처리조세포내Bax표체량증가（P〈0．05），Bcl-2석방량략유강저。상술결과표명：고강도ps맥충통과유도세포조망진이억제료HeLa세포적증식。
In order to study the effects of intense picosecond pulsed electric field (psPEF) on HeLa cells,the psPEF with certain parameters (field intensity of 250 kV/cm, pulse duration of 800 ps, pulse number of 1 000, 3 000, 5 000; repetition frequency of 3 Hz) was investigaed on HeLa cells. Cell growth inhibition was tested by methyl thiazolyl tetrazolium (MTT), changes of concentration of Ca2+ in cells was examined by laser confocal microscopy with cells tabbed with Fluo-3/AM, and release levels of Bax and Bcl-2 were detected by Western blot. Experimental results indicate that cell death is in positive correlation with pulse number, reaching its maximum cell death at 12 h after treatment; significant increases in fluorescence intensity are observed in treated group compared 'with the control group (P〈0.05) by laser eonfocal microscopy, significant increases in Bax are observed in treated group compared with the control group by using western blot {P〈0.05), and slightly decreases in Bcl-2 are observed. The above results demonstrate that proliferation of HeLa cells can be inhibited by psPEF through inducing cell apoptosis.