In-vitro investigation of ALA-induced protoporphyrin IX
In-vitro investigation of ALA-induced protoporphyrin IX
复制标题
DOI：
10.1016/s1011-1344(96)07358-7
复制
发表时间：
1996-11-01
影响因子：
5.4
通讯作者：
Uberriegler, K
中科院分区：
文献类型：
--
作者：
Krammer, B;Uberriegler, K
β-氨基酮丙酸(ALA)诱导的内源性原卟啉IX(PPIX)可用于光动力学杀伤肿瘤细胞。用荧光分光光度法研究了几个参数对人皮肤成纤维细胞PPIX形成的影响。PpIX的形成(1)随ALA的pH值增加，(2)随ALA孵育时间呈适度的S型增加，(3)当ALA浓度达到700微克毫升(-1)时，PPIX的形成增加。其他参数，如细胞清洗程序，对PPIX生产没有影响。ALA的浓度必须比外部原卟啉IX和光敏多糖3高30倍，才能产生相同的细胞毒性损伤。在646 nm处观察到原卟啉的漂白和光产物的产生。通过安装在荧光显微镜上的CCD摄像机获得了关于细胞内PpIX形成动力学及其与细胞结构的拓扑关系的额外信息。在与ALA开始孵育几分钟后，在线粒体中观察到PpIX的生成，随后在细胞质和核膜中重新定位。
Protoporphyrin IX (PpIX) induced endogenously by delta-aminolevulinic acid (ALA), can be used to destroy photodynamically tumor cells. The influence of several parameters on the PpIX formation of human skin fibroblasts was investigated by fluorescence spectrophotometry. The PpIX formation increases (1) with the pH value of ALA, (2) with the ALA incubation time in a moderate sigmoidal manner, and (3) with the ALA concentration up to 700 mu g ml(-1). Other parameters, such as cell washing procedures, have no influence on the PpIX production. ALA has to be applied in a concentration 30 times higher than external protoporphyrin IX and Photosan 3 in order to produce the same cytotoxic damage. Protoporphyrin bleaching and photoproduct generation at 646 nm was observed. Additional information about intracellular PpIX formation kinetics and its topographically correlation to cell structures was gained by a CCD camera mounted on a fluorescence microscope. A few minutes after the onset of incubation with ALA, PpIX generation is observed in the mitochondria, followed by relocalization in the cytoplasm and the nuclear membrane.