CAJ | 학술논문

目的探讨微泡造影剂联合超声辐照对体外培养的人脐静脉血管内皮细胞(HUVEC)膜通透性改变及作用持续时间.方法用连续多普勒超声辐照HUVEC,照射条件为频率1.9 MHz,软组织热指数(TIS)0.8,增益80%,输出声强I SATA为80.0 mW/cm2,照射时间60 s.悬浮HUVEC,加入2%微泡造影剂SonoVue,于照射后0、1、5、30 min、2 h时点分别加入大分子荧光素物质FD500,荧光显微镜下观察FD500进入细胞内的情况,用流式细胞仪测FD500荧光染色阳性的细胞.锥虫蓝染色观察符组样本细胞活性.结果0、1 min组荧光显微镜下大部分HUVEC荧光染色阳性,贴壁后观察可见细胞质内吴现不均质的绿色荧光显影,而细胞核不显影;5、30 min、2 h组仅见少数HUVEC胞质内绿色荧光显影.0、1、5、30 min、2 h组流式细胞仪测荧光染色阳性率分别为(61.63±18.35)%、(57.26±13.53)%、(14.66±6.07)%、(4.12±0.96)%、(5.93±1.45)%,前两组与后者比较,差异有统计学意义(P<0.01).结论微泡造影剂联合诊断级别超声辐照具有增加HUVEC膜通透性的作用,其作用持续时间较为短暂,可能的机制为超声空化导致细胞膜"声穿孔"效应.
목적탐토미포조영제연합초성복조대체외배양적인제정맥혈관내피세포(HUVEC)막통투성개변급작용지속시간.방법용련속다보륵초성복조HUVEC,조사조건위빈솔1.9 MHz,연조직열지수(TIS)0.8,증익80%,수출성강I SATA위80.0 mW/cm2,조사시간60 s.현부HUVEC,가입2%미포조영제SonoVue,우조사후0、1、5、30 min、2 h시점분별가입대분자형광소물질FD500,형광현미경하관찰FD500진입세포내적정황,용류식세포의측FD500형광염색양성적세포.추충람염색관찰부조양본세포활성.결과0、1 min조형광현미경하대부분HUVEC형광염색양성,첩벽후관찰가견세포질내오현불균질적록색형광현영,이세포핵불현영;5、30 min、2 h조부견소수HUVEC포질내록색형광현영.0、1、5、30 min、2 h조류식세포의측형광염색양성솔분별위(61.63±18.35)%、(57.26±13.53)%、(14.66±6.07)%、(4.12±0.96)%、(5.93±1.45)%,전량조여후자비교,차이유통계학의의(P<0.01).결론미포조영제연합진단급별초성복조구유증가HUVEC막통투성적작용,기작용지속시간교위단잠,가능적궤제위초성공화도치세포막"성천공"효응.
Objective To investigate the efficiency of microbubble contrast agents combined with ultrasound exposure on membrane permeability of human umbilical vascular endothelial cells(HUVECs) in vitro. Methods HUVECs were exposed to continuous wave Doppler ultrasound (Frequency, 1.9 MHz;TIS, 0.8 ;Gain ,80% ; I SATA 80.0 mW/cm2) for 1 min, then microbubble contrast agent (2% SonoVue) was added. 0 min, 1 min, 5 min, 30 min and 2 h later, HUVEC suspension was infused with fluorescein isothiocyanate-dextran (FD500) respectively. The uptake of FD500 in HUVEC was observed under fluorescence microscopy. Fluorescent rate of cell was measured with flow cytometry. The survival rate of HUVEC was calculated with trypan blue staining. Results Positive fluoreascence-stain was observed in cytoplasm. The percentages of fluorescence-stained cells were (61.63±18.35)% , (57.26±13.53)%, (14.66±6.07)%, (4.12±0.96)%,(5.93±1.45)% in 0 min, 1 min, 5 min,30 min and 2 h respectively. The percentages of positive fluorescence-stained cells were significantly higher in 0 min, 1 min groups than those of other groups ( P<0.01 ). Conclusions Microbubble contrast agents with diagnostic ultrasound can increase the membrane permeability of HUVEC transitorily. The effect of cavitation can induce cell membrane hole.