原卟啉Ⅸ介导的声动力疗法对S180肿瘤细胞膜损伤研究  

Cell Membrane Damage by Ultrasound in Combination with Protoporphyrin Ⅸ on S180 Tumor Cells

作　　者：薛沁[1] 王筱冰[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：《中国生物医学工程学报》2013年第4期397-402,共6页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金(81000999;10904087)

摘　　要：探讨频率为1.1 MHz、强度为3 W/cm2的聚焦超声,结合1μg/mL的原卟啉Ⅸ(PpⅨ)对S180肿瘤细胞膜结构和功能的损伤作用。超声联合PpⅨ作用于S180肿瘤细胞后,通过酸性磷酸酶活力检测细胞生长活力;扫描电子显微镜观测细胞膜表面超微结构;生化分析方法检测Na+-K+-ATPase、Ca2+-ATPase活性,细胞膜唾液酸(SA)及细胞内谷胱甘肽(GSH)含量;通过荧光酶标仪检测细胞线粒体膜电位及细胞色素C氧化酶含量。实验结果显示:与对照组(Control)、原卟啉Ⅸ组(PpⅨ)、超声组相比,超声联合PpⅨ处理组的细胞生长明显受到抑制,其酸性磷酸酶活力(OD值)由对照组0.63±0.03下降至0.34±0.01;细胞膜结构严重破坏,表现为微绒毛消失和胞质部分成分流失;膜表面重要成分如Na+-K+-ATPase和Ca2+-ATPase活性明显下降(P<0.05),细胞膜唾液酸含量及胞内GSH水平急剧降低(P<0.01)。研究表明:超声联合PpⅨ作用于S180肿瘤细胞后,细胞膜蛋白受到严重破坏,部分膜蛋白流失。研究结果还提示:声动力处理后细胞线粒体膜电位水平显著降低(Rhodamin 123平均荧光强度由对照组14 082.78±840.44下降至7 801.53±409.15);细胞色素C氧化酶活性由对照组的(0.015 6±0.002 0)U/min/mg降低至(0.008±0.000 0)U/min/mg,提示线粒体也可能是超声联合PpⅨ对S180肿瘤细胞的作用靶点。The effects of 1 μg/mL protoporphyrin IX （PpIX）combined with focused ultrasound at the frequency of 1.1 MHz and intensity of 3 W/cm2 on the damage of cell membrane of S180 tumor cell were studied in this paper. Results showed that the ultrasound combined with Pp IX induced more serious damage of the ceils membrane than the ultrasound or the Pp IX alone. We demonstrated that acid phosphatase activity of the treatment group of ultrasound combined with PpIX was decreased from 0. 63 ± 0. 03 to 0. 34 ± 0.01 compared with that of the control group. The structures of cell membrane were destroyed seriously with disappearance of microvilli and partial loss of cytoplasmic constituents in the observation of scanning electron microscopy. Furthermore, after the treatment, the activity of Na -K ＋-ATPase, Ca2 ＋-ATPase, the content of membrane sialic acid （SA） and intracellular GSH were all reduced significantly （P 〈 0.05）. The data indicated that part of membrane proteins was lost. It was also shown that the level of mitoehondrial membrane potential decreased significantly, the mean fluorescence intensity of Rhodamin 123 was decreased from 14 082.78 ± 840.40 to 7 801.53 ± 409.15. And cytochrome C oxidase activity was reduced to 0. 008 U/min/rag. The results indicated that the mitochondria maybe the target of ultrasound PpIX on S180 tumor cells.

关 键 词：聚焦超声 原卟啉Ⅸ 细胞膜 

分 类 号：R318[医药卫生—生物医学工程]