Loh C S, Vernon D, MacRobert A J, Bedwell J, Bown S G, Brown S B
National Medical Laser Centre, University College London Medical School, The Rayne Institute, UK.
J Photochem Photobiol B. 1993 Sep;20(1):47-54. doi: 10.1016/1011-1344(93)80130-2.
The accumulation of endogenous porphyrins in rats following systemic administration of 5-aminolaevulinic acid (ALA) has been examined to assess the photosensitization characteristics of this technique for photodynamic therapy (PDT) and chemical extraction assays with fluorescence and absorbance detection of the porphyrin content have been carried out. We compared the results obtained using quantitative microfluorimetry on normal gastric and colonic tissues in rats at 0.5, 1, 2, 4 and 6 h and chemically induced duodenal tumours 2 and 4.5 h after intravenous administration of ALA at a dose of 200 mg kg-1. With chemical extraction followed by high performance liquid chromatography analysis, protoporphyrin IX (PpIX) was found to be the predominant porphyrin present, reaching peak levels of several microgrammes per gramme at 2-4 h in each type of tissue; a small amount of coproporphyrin was detected at 0.5 and 2 h in normal gastric mucosa and duodenal tumour respectively. Both the extraction assay and quantitative microfluorimetry showed that the porphyrin fluorescence builds up rapidly in the mucosal layers of the colon and stomach, reaching a maximum at 2 h, whereas lower fluorescence levels were found with a slower rate of accumulation in the corresponding muscularis layers. A significant PpIX content was found in the duodenal tumour, with a maximum of 7.1 micrograms g-1 4.5 h after ALA administration. We conclude that systemic administration of ALA can induce effective tissue sensitization with protoporphyrin IX and appears to be a promising technique for PDT.
为评估5-氨基乙酰丙酸(ALA)全身给药后大鼠体内内源性卟啉的蓄积情况,以此来评价该技术用于光动力疗法(PDT)的光敏化特性,并开展了化学提取分析,采用荧光法和吸光法检测卟啉含量。我们比较了在大鼠静脉注射200mg/kg剂量的ALA后0.5、1、2、4和6小时,使用定量显微荧光法在正常胃和结肠组织以及化学诱导的十二指肠肿瘤中2小时和4.5小时获得的结果。通过化学提取后进行高效液相色谱分析,发现原卟啉IX(PpIX)是主要存在的卟啉,在每种组织类型中2-4小时达到每克数微克的峰值水平;在正常胃黏膜和十二指肠肿瘤中分别在0.5小时和2小时检测到少量粪卟啉。提取分析和定量显微荧光法均表明,卟啉荧光在结肠和胃的黏膜层中迅速积累,在2小时达到最大值,而在相应的肌层中荧光水平较低,积累速率较慢。在十二指肠肿瘤中发现了显著的PpIX含量,在ALA给药后4.5小时最高达到7.1μg/g。我们得出结论,ALA全身给药可以诱导原卟啉IX产生有效的组织致敏作用,似乎是一种有前途的PDT技术。