Intra-operative identification of malignant glioma using real-time fluorescence spectroscopic analysis and double staining method

实时荧光光谱分析和双染法术中鉴别恶性胶质瘤

基本信息

批准号：
14571345
负责人：
KUROIWA Toshihiko
金额：
$ 1.6万
依托单位：
Osaka Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

The first purpose of this study is to establish the glioma identification method of PDD using spectroscopic analysis and double staining method. Our second purpose is to develop the next generation PDD agent, which has the excellent tumor specificity. Our third purpose is to achieve the basic studies on the ALA-PDT.To analyze the fluorescence spectrum using fiber-optic spectroscopic tumor sensor, we could identify the tumor specific spectrum. The time resolution of spectroscopic identification is very short, so that this sensing method can be adapted for the sensor type surgical robot system. Next, we developed the new fluorescent agent (FLS-HAS) to conjugate the FLS and human albumin. The FLS-HAS has the excellent tumor specificity compared with 5-ALA. We found that the EPR (electron spin resonance method) could be applicable to the direct detection of ROSs (reactive oxygen species) in the living cell during PDT. The major ROSs of ALA-PDT was singlet oxygen. As for the mechanism of cell death after ALA-PDT, we found that the increases in the activity of both caspase-3 and -9, a decrease in MMP (mitochondrial membrane potential), and a marked increase in cytochrome c in the cytosolic fl-action were found after cells were subjected to PDT. These results indicate that a dysfunction of MMP is followed by mitochondrial cytochrome c release, which triggers apoptosis through a mitochondrial pathway.

这项研究的第一个目的是使用光谱分析和双重染色方法建立PDD的神经胶质瘤鉴定方法。我们的第二个目的是开发下一代PDD剂，该PDD剂具有出色的肿瘤特异性。我们的第三目的是实现ALA-PDT的基础研究。使用光纤光谱肿瘤传感器分析荧光光谱，我们可以鉴定肿瘤特异性光谱。光谱识别的时间分辨率非常短，因此可以为传感器类型的手术机器人系统调整这种传感方法。接下来，我们开发了新的荧光剂（FLS-HAS）来结合FLS和人白蛋白。与5-ALA相比，FLS-HAS具有出色的肿瘤特异性。我们发现，EPR（电子自旋共振方法）可用于PDT期间活细胞中Ross（反应性氧）的直接检测。 ALA-PDT的主要Ross是单线氧。至于ALA-PDT后细胞死亡的机制，我们发现CASPASE-3和-9的活性增加，MMP的降低（线粒体膜电位），并且在细胞后发现细胞溶剂中的细胞色素C显着增加，使细胞受到PDT的影响。这些结果表明，MMP的功能障碍之后是线粒体细胞色素C释放，这会通过线粒体途径触发凋亡。