Real-time visualization of photochemically induced fluorescence of 8-halogenated quinolones: lomefloxacin, clinafloxacin and Bay3118 in live human HaCaT keratinocytes.
Real-time visualization of photochemically induced fluorescence of 8-halogenated quinolones: lomefloxacin, clinafloxacin and Bay3118 in live human HaCaT keratinocytes.
复制标题
活人 HaCaT 角质形成细胞中 8-卤代喹诺酮类药物洛美沙星、克那沙星和 Bay3118 光化学诱导荧光的实时可视化。
DOI：
10.1111/j.1751-1097.2010.00741.x
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发表时间：
2010
影响因子：
3.3
通讯作者：
He,Yu-Ying
中科院分区：
文献类型：
--
作者：
Koker,EdmondB;Bilski,PiotrJ;Motten,AnnG;Zhao,Baozhong;Chignell,ColinF;He,Yu-Ying
卤代喹诺酮类药物是一种有效且广泛使用的抗微生物药物，可阻断微生物DNA合成。然而，它们通过吸收紫外线诱导不良光反应，包括光毒性和光致癌性。光毒性反应可能是单线态氧的光敏化、自由基的产生和/或由光脱卤产生的其他活性物质的结果。在这里，我们报告使用激光扫描共聚焦显微镜检测和跟踪荧光变化的一个单卤代和三个二卤代喹诺酮类药物在活的人表皮角质形成细胞在原位共聚焦激光照射过程中，在真实的时间。荧光图像分析和与LysoTracker探针共染色表明，溶酶体是药物定位和光转化的优先位点。由于溶酶体环境是相对酸性的，我们还确定了低pH值如何影响脱卤和伴随的荧光。随着紫外线照射的持续，BAY y3118和克林沙星的光产物的荧光增加，而洛美沙星和阿托沙星的荧光减少。我们的图像不仅有助于将这些光毒性剂定位在细胞中，而且还提供了动态监测其在细胞环境中的光转化的手段。
Halogenoquinolones are potent and widely used antimicrobials blocking microbial DNA synthesis. However, they induce adverse photoresponses through the absorption of UV light, including phototoxicity and photocarcinogenicity. The phototoxic responses may be the result of photosensitization of singlet oxygen, production of free radicals and/or other reactive species resulting from photodehalogenation. Here, we report the use of laser scanning confocal microscopy to detect and to follow the fluorescence changes of one monohalogenated and three di‐halogenated quinolones in live human epidermal keratinocyte cells duringin situirradiation by confocal laser in real time. Fluorescence image analysis and co‐staining with the LysoTracker probe showed that lysosomes are a preferential site of drug localization and phototransformations. As the lysosomal environment is relatively acidic, we also determined how low pH may affect the dehalogenation and concomitant fluorescence. With continued UV irradiation, fluorescence increased in the photoproducts from BAY y3118 and clinafloxacin, whereas it decreased for lomefloxacin and moxifloxacin. Our images not only help to localize these phototoxic agents in the cell, but also provide means for dynamic monitoring of their phototransformations in the cellular environment.