1 Division of Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
2 Laboratory of Physical Chemistry, Showa Pharmaceutical University, Machida, Tokyo, Japan.
Mol Imaging. 2019 Jan-Dec;18:1536012118820421. doi: 10.1177/1536012118820421.
Oxidative stress plays an important role in the onset of many neuronal and peripheral disorders. We examined the feasibility of obtaining semiquantitative fluorescent images of reactive oxygen species (ROS) generation in mouse brain and kidney utilizing a planar laser scanner and dihydroethidium (DHE).
To investigate ROS generation in brain, sodium nitroprusside was injected into the striatum. Dihydroethidium was injected into the tail vein. After DHE injection, tissue slices were analyzed utilizing a planar laser scanner. For kidney study, cis-diamminedichloroplatinum [II] (cisplatin) was intraperitoneally administrated into mice.
Clear and semiquantitative fluorescent images of ROS generation in the mouse brain and kidney were obtained. Furthermore, the fluorescence intensity was stable and not affected by fading. Sodium nitroprusside induced approximately 6 times the fluorescence accumulation in the brain. Cisplatin caused renal injury in all mice, and in comparison with control mice, more than 10 times fluorescence accumulation was observed in the renal medulla with tubular necrosis and vacuolization.
We successfully obtained ex vivo semiquantitative fluorescent images of ROS generation utilizing a planar laser scanner and DHE. This simple method is useful for ROS detection in several ROS-related animal models and would be applicable to a variety of biochemical processes.
氧化应激在许多神经元和外周疾病的发生中起着重要作用。我们利用平面激光扫描仪和二氢乙锭（DHE）检查了在小鼠脑和肾中获得活性氧（ROS）生成的半定量荧光图像的可行性。
为了研究脑内 ROS 的生成，将硝普钠注射到纹状体中。将二氢乙锭注入尾静脉。DHE 注射后，利用平面激光扫描仪分析组织切片。对于肾脏研究，顺铂[II]（顺铂）被腹膜内给予小鼠。
在小鼠脑和肾中获得了清晰且半定量的 ROS 生成荧光图像。此外，荧光强度稳定，不受褪色影响。硝普钠诱导脑中的荧光积累约增加 6 倍。顺铂导致所有小鼠的肾脏损伤，与对照组小鼠相比，在肾小管坏死和空泡化的肾髓质中观察到的荧光积累超过 10 倍。
我们成功地利用平面激光扫描仪和 DHE 获得了 ROS 生成的离体半定量荧光图像。这种简单的方法可用于几种与 ROS 相关的动物模型中的 ROS 检测，并且适用于多种生化过程。
