聚焦超声对艾氏腹水瘤细胞膜损伤作用的研究  

Damaging the membrane of Ehrlich ascitic tumor cells with focused ultrasound

作　　者：郝巧[1] 王筱冰[1] 仝万艳[1] 李晨笛[1] 刘全宏[1] 

机构地区：[1]陕西师范大学生命科学学院,西安710062

出　　处：《中华物理医学与康复杂志》2009年第1期16-20,共5页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金

摘　　要：目的 初步探讨频率为2.2 MHz的聚焦超声对艾氏腹水瘤(EAC)细胞膜的损伤作用.方法 台盼蓝拒染法检测不同强度聚焦超声辐射下的细胞的相对存活率;扫描电镜观察细胞膜表面超微结构的变化;流式细胞仪检测辐射后大分子异硫氰酸荧光素钠标记的右旋糖酐(FD500)进入细胞内的量;乳酸脱氢酶(LDH)测定试剂盒检测不同辐射后细胞悬液中乳酸脱氢酶的释放量.结果 扫描电镜下观察到EAC细胞膜表面具有声孔现象;聚焦超声辐射30 s与60 s组细胞胞内FD500含量分别为11.46%和18.50%,较对照组含量(0.21%)明显增加;而聚焦超声30 s组和60 s组胞外LDH的活性分别为(2.94±0.02)U/L、(3.28±0.04)U/L,即LDH释放量均随辐射时间的延长而增加.结论 聚焦超声辐射可导致细胞膜通透性改变,其损伤程度与聚焦超声辐射时间有一定的相关性.Objective To study the damage focused ultrasound inflicts on the membrane permeability of Ehrlich ascitic tumor (EAC) cells and the relationship between changes in membrane permeability and focused ultra-sound exposure time. Methods The relative survival rate of tumor cells was examined at various intensities and dif-ferent exposure times using focused 2.2 MHz ultrasound. The uhrastructure changes were evaluated with a scanning electron microscope after different exposures. Membrane permeability was investigated by incorporating fluorescein isothiocyanate dextran (FD5OO) , and membrane damage was evaluated by measuring lactate dehydrogenase (LDH) release. Results Morphological observation showed there were numerous microvilli on the surface of un-exposed cells. When the cells had been irradiated with focused ultrasound for 30 s there was only a slight effect on the shape of the cells and the number of microvilli was slightly reduced. When the cells were exposed to ultrasound for 60 s, the surface of many cells became relatively smooth with no obvious microvilli, and several small craters were seen on the surfaces of cells where the cytoplasm seemed to have extruded through the membrane. The cell membrane was seri-ously damaged by sonoporation. The loading of FD500 in the unexposed cells was only 0.21%. When the cells had been sonicated with focused ultrasound for 30 s or 60 s, the loading of FD500 increased to 11.46% and 18.50% re-spectively. The released LDH activities in the 30 s group and 60 s group were 2.94±0.02 and 3.28±0.04 U/L, respectively. The activities of LDH increaased as the focused ultrasound exposure time was prolonged. Conclusion Focused ultrasound may damage the cell membrane permeability of EAC cells, and the damage increases as the expo-sure time is prolonged from 30 s to 60 s.

关 键 词：聚焦超声 声孔效应 细胞膜 

分 类 号：R73-3[医药卫生—肿瘤]