HMME介导的光动力对兔血管平滑肌细胞生长抑制及诱导细胞凋亡的作用  

The Cytotoxicity and Apoptosis Induced by HMME-based Photodynamic Therapy in Rabbit Vascular Smooth Muscle Cells

作　　者：阴慧娟[1] 李晓原[1] 林红[2] 刘建中[3] 于红奎[2] 李燕[4] 

机构地区：[1]中山大学基础医学院生物医学工程系激光医学研究室,广东广州510080 [2]中山大学附属第一医院,广东广州510080 [3]中山大学基础医学院生物学教研室,广东广州510080 [4]中山大学基础医学院教学实验中心,广东广州510080

出　　处：《中国医学物理学杂志》2006年第5期341-344,共4页Chinese Journal of Medical Physics

基　　金：广东省医学科研基金项目(A2006173)

摘　　要：目的:观察原代培养的兔血管平滑肌细胞在HMME-PDT作用下生长的变化及其死亡方式。方法:常规台盼兰染色观察HMME-PDT作用后6h和24 h对细胞的杀伤作用,HMME的浓度为10μg/ml,激光剂量为2.4～9.6 J/cm2;HE染色法观察PDT后24 h细胞形态的变化;流式细胞仪AnnexinⅤ/PI双染法检测细胞死亡方式;共聚焦显微镜观察HMME在线粒体的定位。结果:台盼兰法检测结果显示随着PDT能量密度的增加细胞的存活率逐渐下降,光镜下可见大部分细胞呈死亡或凋亡样改变,AnnexinⅤ/PI法检测显示PDT 24 h后凋亡率可达50.5%,共聚焦显微镜观察到HMME在线粒体内有分布。结论:HMME-PDT能显著抑制兔血管平滑肌细胞的生长,并使其发生明显的凋亡。Objective： To observe the change of rabbit vascular smooth muscle cells＇ proliferation and the type of death induced by HMME-PDT. Methods： The cytotoxic effect of HMME-PDT on rabbit vascular smooth muscle cells was studied by Trypan Blue assay, HMME at 10μg/ml concentration and the light dose at 2.4-9.6 J/cm^2 were selected in the studies. The morphological character 24h post-PDT was investigated by HE. Annexin V and propidium iodide （PI） binding assays were performed to analyze the characteristics of cell death after HMME-PDT. Furthermore, The intracellular distributions of the HMME were measured by the confocal laser scanning microscope. Results： It was showed the photocytotoxity to VSMC cells was dose related by Trypan Blue assay. Histology observing suggested HMME-PDT could induce cell death through apoptosis or necrosis, and the apoptosic rate was up to 50.5% by Annexin V/PI assay. Moreover, the fluorescence images of HMME intracellular localization demonstrated that the HMME diffused into the mitochondria. Conclusions： HMME-PDT could significantly inhibite VSMC proliferation and induce apoptosis.

关 键 词：血卟啉单甲醚 光动力 血管平滑肌细胞 凋亡 

分 类 号：R318.51[医药卫生—生物医学工程]