原卟啉Ⅸ介导的声动力疗法对S180 肿瘤细胞膜损伤研究

doi:10.3969/j.issn.0258-8021. 2013. 04.03

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摘要探讨频率为1.1 MHz、强度为3 W/cm2的聚焦超声，结合1 μg/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。

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	薛沁王筱冰^*刘全宏

关键词 ：聚焦超声, 原卟啉Ⅸ, 细胞膜

Abstract：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/cm2 on the damage of cell membrane of S180 tumor cell were studied in this paper. Results showed that the ultrasound combined with PpⅨ induced more serious damage of the cells membrane than the ultrasound or the PpⅨ 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, Ca^2+-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 mitochondrial 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/mg. The results indicated that the mitochondria maybe the target of ultrasound PpⅨ on S180 tumor cells.

Key words： focused ultrasound frotoporphyrin Ⅸ cell membrane

基金资助:国家自然科学基金（81000999，10904087）

引用本文:

薛沁王筱冰^*刘全宏. 原卟啉Ⅸ介导的声动力疗法对S180 肿瘤细胞膜损伤研究[J]. 中国生物医学工程学报, 2013, 32(4): 397-402.
XUE Qin WANG Xiao Bing^* LIU Quan Hong. Cell Membrane Damage by Ultrasound in Combination with Protoporphyrin Ⅸ on S180 Tumor Cells. journal1, 2013, 32(4): 397-402.

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http://cjbme.csbme.org/CN/10.3969/j.issn.0258-8021. 2013. 04.03 或 http://cjbme.csbme.org/CN/Y2013/V32/I4/397

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