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首页> 外文期刊>Romanian Journal of Biophysics >ALA UPTAKE AND LASER INDUCED ROS PRODUCTION IN KERATINOCYTES
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ALA UPTAKE AND LASER INDUCED ROS PRODUCTION IN KERATINOCYTES

机译：角质形成细胞中ALA摄取和激光诱导的ROS产生

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Our study aims to assess the effect of photodynamic therapy (PDT) using ALA (5-aminolaevulinic acid, a precursor of intracellular porphyrins) and red LASER light on Human Caucasian dysplastic oral keratinocytes (DOK). As a first step we determined the best experimental conditions and the amount of reactive oxygen species (ROS) produced as a result of ALA-PDT. DOK cells were incubated with various concentrations of ALA, for different periods of time, in order to determine the optimal concentration of the photosensitizer and duration of incubation. The intracellular amount of ALA-induced protoporphyrin IX (PpIX) was estimated by its fluorescence emission spectra. The photolysis of PpIX in cells irradiated with a red LASER was measured by a decrease in the intensity of these spectra. Intracellular ROS were evaluated using a fluorescent nonselective marker, 2’,7’–dichlorofluorescein diacetate (DCFDA). Results: The ALA concentration corresponding to optimal cell uptake and PpIX production was determined to be 1mM, over a 24h incubation period. The photolysis of PpIX in cells irradiated with red LASER light was proven by the decrease of PpIX emission peak. The rate of ROS concentration increase (as determined with DCFDA) was diminished by adding vitamin C in the cellular suspension.
机译：我们的研究旨在评估使用ALA（5-氨基乙酰戊酸，细胞内卟啉的前体）和红色LASER光对人高加索发育异常的口腔角质形成细胞（DOK）进行光动力疗法（PDT）的效果。第一步，我们确定了最佳实验条件以及ALA-PDT产生的活性氧（ROS）的量。将DOK细胞与各种浓度的ALA孵育不同的时间，以确定光敏剂的最佳浓度和孵育时间。通过其荧光发射光谱估算ALA诱导的原卟啉IX（PpIX）的细胞内量。通过降低这些光谱的强度来测量在用红色激光照射的细胞中PpIX的光解作用。使用荧光非选择性标记2'，7'-二氯荧光素二乙酸酯（DCFDA）评估细胞内ROS。结果：在24小时的孵育时间内，对应于最佳细胞摄取和PpIX产生的ALA浓度确定为1mM。 PpIX发射峰的减少证明了PpIX在红色激光激光照射的细胞中的光解作用。通过在细胞悬浮液中添加维生素C，可以减少ROS浓度增加的速率（由DCFDA确定）。

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