基于ALA-PDT中PpIX亚细胞分布位点光漂白实验研究

Experimental Research on Subcellular Distribution of PpIX Photobleaching in HL60 Based on ALA-PDT

摘要:

随着光动力学疗法(photodynamic therapy，PDT)基础研究的不断深入和临床应用的广泛开展，光敏剂的光漂白现象日渐成为PDT研究的热点问题。实验在ALA的终浓度为10 mM/ml，辐射光波长为410 nm，辐照光功率为5 mW/cm2等条件下采用荧光探针标记技术在自行设计的光动力疗法反应室进行了PDT实验研究。Cell Counting Kit-8 (CCK-8)法用来检测细胞的活性，通过获得的荧光光谱研究了亚细胞的光漂白情况。结果表明：以每种细胞器光照前的平均荧光强度为基准，则光照60 min后线粒体、溶酶体、内质网的荧光强度下分别降低了95%、91.5%和51.2%。线粒体区域光漂白速率较快，其荧光强度的变化基本表征了HL60细胞活性的变化，推断线粒体是光动力效应的主要作用靶点之一。

Abstract: With the development of the fundamental research of photodynamic therapy (PDT) and its clinical applica-tion, the photobleaching properties of photosensitizers have become a hotspot of research. The experimental research of ALA mediated PDT was carried out in the own designed reaction chamber when the final concentration of ALA is 10 mM/ml, the wavelength and the output power of the light source are 410 nm and 5 mW/cm2 separately. Cell Counting Kit-8 (CCK-8) was used to evaluate the cell viability while the photobleaching of PpIX in the subcellular structure of HL60 was detected by fluorescence spectroscopy. The results showed that after 60 min irradiation the fluorescence in-tensity of mitochondria, lysosomes and endoplasmic reticulum had decreased by 95%, 91.5% and 51.2% respectively compared to that before irradiation. The rate of photobleaching in mitochondria which can characterize the activity of HL60 was significantly higher than others, this suggest that mitochondria might be one of main therapeutic targets of photodynamic therapy.