Real-time monitoring of oxidative stress in live mouse skin.
Real-time monitoring of oxidative stress in live mouse skin.
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DOI：
10.1038/jid.2013.428
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发表时间：
2014-06
期刊：
影响因子：
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通讯作者：
Alexander M. Wolf;K. Nishimaki;Naomi Kamimura;S. Ohta
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文献类型：
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作者：
Alexander M. Wolf;K. Nishimaki;Naomi Kamimura;S. Ohta
氧化应激与许多与年龄相关的疾病以及衰老过程本身有关。由于缺乏可用于活体动物的灵敏检测方法，减少氧化应激的干预措施的发展受到阻碍。我们产生的转基因小鼠表达比例氧化还原敏感的绿色荧光蛋白（roGFP）的细胞质或线粒体的几个组织，包括皮肤表皮角质形成细胞。杂交成无毛白化病小鼠允许皮肤氧化状态的非侵入性光学测量。局部应用过氧化氢乳剂在几分钟内将角质形成细胞的氧化还原状态向氧化方向转变，并且可以通过荧光比率成像进行真实的实时观察。将皮肤暴露于365 nm UVA辐射氧化定位在角质形成细胞线粒体中的roGFP，但当roGFP定位在胞质溶胶中时不氧化。这表明介导UVA诱导的氧化应激的大量内源性光敏剂位于线粒体中。紫外线是皮肤老化的主要环境原因，UVA介导的氧化应激与人类皱纹的发展有关。直接测量活体动物的确定细胞室中的氧化还原状态应该是用于评估旨在调节氧化应激的治疗的有力且方便的工具。
Oxidative stress is involved in many age-associated diseases, as well as in the aging process itself. The development of interventions to reduce oxidative stress is hampered by the absence of sensitive detection methods that can be used in live animals. We generated transgenic mice expressing ratiometric redox-sensitive green fluorescent protein (roGFP) in the cytosol or mitochondria of several tissues, including skin epidermal keratinocytes. Crossbreeding into hairless albino mice allowed noninvasive optical measurement of skin oxidative state. Topical application of hydrogen peroxide emulsion shifted the keratinocyte redox state toward oxidation within minutes and could be observed in real time by fluorescence ratio imaging. Exposing skin to 365 nm UVA radiation oxidized roGFP localized in keratinocyte mitochondria, but not when roGFP was localized in the cytosol. This suggests that significant amounts of the endogenous photosensitizers that mediate UVA–induced oxidative stress are located in the mitochondria. UVR is the major environmental cause of skin aging and UVA–mediated oxidative stress has been associated with the development of wrinkles in humans. Direct measurements of redox state in defined cell compartments of live animals should be a powerful and convenient tool for evaluating treatments that aim to modulate oxidative stress.