Monitoring singlet oxygen and hydroxyl radical formation with fluorescent probes during photodynamic therapy.
Monitoring singlet oxygen and hydroxyl radical formation with fluorescent probes during photodynamic therapy.
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DOI：
10.1111/j.1751-1097.2009.00555.x
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发表时间：
2009-09
影响因子：
3.3
通讯作者：
Kessel D
中科院分区：
文献类型：
--
作者：
Price M;Reiners JJ;Santiago AM;Kessel D
单线态氧 (1O2) 是光动力疗法 (PDT) 方案中产生的主要氧化剂，涉及导致 II 型反应的敏化剂。 1O2 可以产生额外的活性氧（ROS），例如羟基自由基（·OH）。目前的研究旨在评估 3′-p-（氨基苯基）荧光素（APF）和 3′-p-（羟苯基）荧光素（HPF）作为在 PDT 相关条件下检测 1O2 和·OH 的探针。无细胞研究表明，APF 和 HPF 暴露于水溶性光敏剂 (TPPS) 照射产生的 1O2 后都会转化为荧光产物，并且 APF 的灵敏度是 HPF 的 35 倍。使用 1O2 探针单线态氧传感器绿色 (SOSG)，我们证实 1 mm NaN3 猝灭 1O2 诱导的 APF /HPF 荧光，而 1% DMSO 没有影响。 APF 和 HPF 在无细胞系统中通过芬顿反应与 H2O2 生成的·OH 相互作用时也产生荧光产物。 DMSO 在剂量低至 0.02% 时即可淬灭 APF /HPF 和·OH 之间的荧光相互作用。尽管NaN3 预计会淬灭•OH 诱导的APF /HPF 荧光，但在•OH 存在的情况下，将NaN3 与APF 或HPF 共孵育会显着增强荧光。用苯并卟啉衍生物光敏的培养的L1210细胞在照射后立即表现出APF荧光。加入 DMSO 或铁螯合剂去铁胺可抑制大约 50% 的细胞荧光。后两种药物的组合并没有增强抑制作用。我们得出的结论是，如果分别在存在和不存在 DMSO 的情况下进行研究，APF 可用于监测接受 PDT 的细胞中 1O2 和·OH 的形成。被 DMSO 猝灭的那部分荧光将代表·OH 的贡献。该程序可以代表一种在 PDT 背景下评估两种 ROS 形成的有用方法。
Singlet oxygen (1O2) is the primary oxidant generated in photodynamic therapy (PDT) protocols involving sensitizers resulting in type II reactions. 1O2 can give rise to additional reactive oxygen species (ROS) such as the hydroxyl radical (•OH). The current study was designed to assess 3′-p-(aminophenyl) fluorescein (APF) and 3′-p-(hydroxyphenyl) fluorescein (HPF) as probes for the detection of 1O2 and •OH under conditions relevant to PDT. Cell-free studies indicated that both APF and HPF were converted to fluorescent products following exposure to 1O2 generated by irradiation of a water-soluble photosensitizing agent (TPPS) and that APF was 35-fold more sensitive than HPF. Using the 1O2 probe singlet oxygen sensor green (SOSG) we confirmed that 1 mm NaN3 quenched 1O2-induced APF /HPF fluorescence, while 1% DMSO had no effect. APF and HPF also yielded a fluorescent product upon interacting with •OH generated from H2O2 via the Fenton reaction in a cell-free system. DMSO quenched the fluorogenic interaction between APF /HPF and •OH at doses as low as 0.02%. Although NaN3 was expected to quench •OH-induced APF /HPF fluorescence, co-incubating NaN3 with APF or HPF in the presence of •OH markedly enhanced fluorescence. Cultured L1210 cells that had been photosensitized with benzoporphyhrin derivative exhibited APF fluorescence immediately following irradiation. Approximately 50% of the cellular fluorescence could be suppressed by inclusion of either DMSO or the iron-chelator desferroxamine. Combining the latter two agents did not enhance suppression. We conclude that APF can be used to monitor the formation of both 1O2 and •OH in cells subjected to PDT if studies are performed in the presence and absence of DMSO, respectively. That portion of the fluorescence quenched by DMSO will represent the contribution of •OH. This procedure could represent a useful means for evaluating formation of both ROS in the context of PDT.
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影响因子：
4.8
作者：
Setsukinai, K;Urano, Y;Nagano, T
通讯作者：
Nagano, T
影响因子：
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通讯作者：
HALLIWELL, B