The detection and quantification of highly reactive oxygen species using the novel HPF fluorescence probe in a rat model of focal cerebral ischemia
The detection and quantification of highly reactive oxygen species using the novel HPF fluorescence probe in a rat model of focal cerebral ischemia
复制标题
DOI：
10.1016/j.neures.2005.08.002
复制
发表时间：
2005-11-01
影响因子：
2.9
通讯作者：
Saito, N
中科院分区：
文献类型：
--
作者：
Tomizawa, S;Imai, H;Saito, N
采用新型荧光探针2-[6-（4'-羟基）苯氧基- 3h -杂原-3-on-9-基]苯甲酸（HPF）研究了体外和体内缺血条件下高活性氧（hROS）的生成。在体外实验中，我们用ht22细胞证实，ht22主要在细胞质中检测到，这与线粒体的位置一致，在谷氨酸诱导的氧化应激作用下，其HPF荧光从6 h到24 h逐渐增加。在体内实验中，对大鼠进行了永久性和暂时性大脑中动脉闭塞（MCAO）的诱导。脑切片在含HPF的人工培养基中孵育。在永久性和暂时性MCAO模型中，MCAO后4 h，缺血核心和梗死周围区域均存在HPF荧光增强区域。该区域超出了缺血损伤的边界，进入生化活性组织。瞬时MCAO增强的荧光强度高于永久NICAO模型。羟基自由基清除剂MCI-186显著抑制荧光增强。本研究表明，HPF在局灶性脑缺血和氧化应激细胞模型中检测hROS具有很高的敏感性和特异性。(c) 2005爱思唯尔爱尔兰有限公司和日本神经科学学会。版权所有。
A novel fluorescence probe, 2-[6-(4'-hydroxy) phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (HPF) was used to investigate the generation of highly reactive oxygen species (hROS) under ischemia both in vitro and in vivo. In the in vitro study, HT 22 cells were used to demonstrate that was predominantly detected in the cytoplasm, which coincides with the location of the mitochondria and then its HPF fluorescence gradually increased from 6 to 24 h due to glutamate induced oxidative stress. In the in vivo study, the permanent and transient middle cerebral artery occlusion (MCAO) was induced in rats. Brain slices were incubated in an artificial medium containing HPF. The area of enhanced HPF fluorescence existed in both the ischemic core and the peri-infarct area at 4 h after MCAO in both permanent and transient MCAO models. The area extended beyond the boundary of the ischemic damage into biochemically viable tissue. The enhanced fluorescent intensity following transient MCAO was higher than that observed in the permanent NICAO model. Hydroxyl radical scavenger, MCI-186 significantly suppressed the enhanced fluorescence intensity. This study demonstrated that HPF has a high sensitivity and specificity for the detection of hROS in focal cerebral ischemia as well as in a cellular model of oxidative stress. (c) 2005 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.