Yasui H, Sakurai H
Department of Analytical and Bioinorganic Chemistry, Kyoto Pharmaceutical University, 5 Nakauchi-cho, Misasagi, Yamashina-ku, Kyoto, 607-8414, Japan.
Biochem Biophys Res Commun. 2000 Mar 5;269(1):131-6. doi: 10.1006/bbrc.2000.2254.
The recent increase of ultraviolet (UV) rays on Earth due to the increasing size of the ozone hole is suggested to be harmful to life and to accelerate premature photoaging of the skin. The detrimental effects of UV radiation on the skin are associated with the generation of reactive oxygen species (ROS) such as superoxide anion radical (*O(-)(2)), hydrogen peroxide (H(2)O(2)), hydroxyl radical (*OH), and singlet oxygen ((1)O(2)). However, direct proof of such ROS produced in the skin under UV irradiation has been elusive. In this study, we report first in vivo detection and imaging of the generated ROS in the skin of live mice following UVA irradiation, in which both a sensitive and specific chemiluminescence probe (CLA) and an ultralow-light-imaging apparatus with a CCD camera were used. In addition, we found that *O(-)(2) is formed spontaneously and (1)O(2) is generated in the UVA-irradiated skin. This method should be useful not only for noninvasive investigation of the spatial distribution and quantitative determination of ROS in the skin of live animals, but also for in vivo evaluation of the protective ability of free radical scavengers and antioxidants.
由于臭氧层空洞不断扩大，地球上紫外线（UV）辐射量最近有所增加，这被认为对生命有害，并会加速皮肤过早出现光老化。紫外线辐射对皮肤的有害影响与活性氧（ROS）的产生有关，如超氧阴离子自由基（*O(-)(2)）、过氧化氢（H(2)O(2)）、羟基自由基（OH）和单线态氧（(1)O(2)）。然而，紫外线照射下皮肤中产生此类活性氧的直接证据一直难以获得。在本研究中，我们首次报告了在长波紫外线（UVA）照射后，对活体小鼠皮肤中产生的活性氧进行体内检测和成像，其中使用了一种灵敏且特异的化学发光探针（CLA）和一台配备电荷耦合器件（CCD）相机的超低光成像设备。此外，我们发现O(-)(2)在UVA照射的皮肤中自发形成，且会产生(1)O(2)。该方法不仅应有助于对活体动物皮肤中活性氧的空间分布进行无创研究和定量测定，还可用于体内评估自由基清除剂和抗氧化剂的保护能力。
