微泡造影剂联合超声辐照短暂性增加血管内皮细胞膜通透性  

Transitory membrane permeability increase of human umbilical vascular endothelial cells by microbubble contrast agents combined with ultrasound exposure

作　　者：汤庆[1] 何小洪[2] 袁敏翰[1] 周大治[1] 陈毓菁[1] 何练图[1] 

机构地区：[1]广州医学院附属第一医院超声科,510120 [2]中山大学附属第一医院心内科实验室

出　　处：《中华生物医学工程杂志》2008年第3期218-220,共3页Chinese Journal of Biomedical Engineering

基　　金：广东省自然科学基金（0h6301115）

摘　　要：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1．9MHz，软组织热指数（TJS）0．8，增益80％，输出声强ISATA为80．0mW／cm^2，照射时间60s。悬浮HUVEC，加入2％微泡造影剂SonoVue，于照射后0、1、5、30min、2h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。锥虫蓝染色观察各组样本细胞活性。结果0.1min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呈现不均质的绿色荧光显影，而细胞核不显影；5、30min、2h组仅见少数HUVEC胞质内绿色荧光显影。0、1、5、30min、2h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”效虚。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 roW/era2） for 1 rain, then mierobubble contrast agent （2% SonoVue）was added. 0 rain, 1 rain, 5 rain, 30 rain and 2 h later, HUVEC suspension was infused with fluorescein isothioeyanate-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 rain,5 rain,30 min and 2 h respectively. The percentages of positive fluorescence-stained ceils were significantly higher in 0 rain, 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.

关 键 词：超声空化效应 微泡造影剂 人脐静脉血管内皮细胞 膜通透性 

分 类 号：R445.1[医药卫生—影像医学与核医学]