Real-time monitoring of superoxide accumulation and antioxidant activity in a brain slice model using an electrochemical cytochrome c biosensor.
Real-time monitoring of superoxide accumulation and antioxidant activity in a brain slice model using an electrochemical cytochrome c biosensor.
复制标题
DOI：
10.1016/j.freeradbiomed.2012.10.540
复制
发表时间：
2012-12-15
影响因子：
7.4
通讯作者：
Andreescu, Silvana
中科院分区：
文献类型：
--
作者：
Ganesana, Mallikarjunarao;Erlichman, Joseph S.;Andreescu, Silvana
关键词：
活性氧簇的过度产生和由此产生的损害是许多疾病的病理学的核心。由于缺乏能够连续测量的特异性探针和技术，对活性氧的时空积累的研究受到限制。我们证明了使用一个小型化的电化学细胞色素C（Cyt C）生物传感器的实时测量和定量评估超氧化物的生产和灭活的天然和工程抗氧化剂在急性制备的小鼠脑切片。在对照条件下，400 μm厚切片中大脑海马区产生的超氧自由基完全在电极的检测范围内。将切片暴露于缺血条件下使超氧化物产生增加两倍，并且切片的测量值在3-4小时内稳定。芪衍生物和阴离子通道抑制剂4，4 ′-二异硫氰基-2，2 ′-二磺酸芪（DIDS）在对照条件下显著降低细胞外超氧化物信号，表明超氧化物跨膜通量进入细胞外空间可能是正常氧化还原信号的一部分。超氧化物歧化酶（SOD），以剂量依赖性的方式减少超氧化物信号的外源性添加在海马细胞释放的超氧化物的电极的特异性进行了验证。添加SOD模拟物、氧化铈纳米颗粒（纳米二氧化铈）也观察到类似的结果，其中平均直径为15 nm的纳米二氧化铈的超氧阴离子自由基清除活性相当于每添加1 μg/ml纳米二氧化铈527 U的SOD。这项研究表明，电化学生物传感器的潜力，用于研究实时动态的活性氧物种在生物模型和这些测量的效用，在定义的相对贡献超氧化物氧化损伤。
The overproduction of reactive oxygen species and resulting damage are central to the pathology of many diseases. The study of the temporal and spatial accumulation of reactive oxygen species has been limited due to the lack of specific probes and techniques capable of continuous measurement. We demonstrate the use of a miniaturized electrochemical cytochrome C (Cyt C) biosensor for real-time measurements and quantitative assessment of superoxide production and inactivation by natural and engineered antioxidants in acutely prepared brain slices from mice. During control conditions, superoxide radicals produced from the hippocampal region of the brain in 400 μm thick sections were well within the range of detection of the electrode. Exposure of the slices to ischemic conditions increased the superoxide production two fold and measurements from the slices were stable over a 3–4 hour period. The stilbene derivative and anion channel inhibitor, 4,4′-diisothiocyano-2,2′-disulfonic stilbene (DIDS), markedly reduced the extracellular superoxide signal under control conditions suggesting that a transmembrane flux of superoxide into the extracellular space may occur as part of normal redox signaling. The specificity of the electrode for superoxide released by cells in the hippocampus was verified by the exogenous addition of superoxide dismutase (SOD) which decreased the superoxide signal in a dose-dependent manner. Similar results were seen with the addition of the SOD-mimetic, cerium oxide nanoparticles (nanoceria) where the superoxide anion radical scavenging activity of nanoceria with an average diameter of 15 nm was equivalent to 527 U of SOD for each 1 μg/ml of nanoceria added. This study demonstrates the potential of electrochemical biosensors for studying real-time dynamics of reactive oxygen species in a biological model and the utility of these measurements in defining the relative contribution of superoxide to oxidative injury.
登录
查看更多内容
影响因子：
4.8
作者：
Bahamonde, MI;Fernández-Fernández, JM;Valverde, MA
通讯作者：
Valverde, MA
影响因子：
11.2
作者：
FLOYD, RA;CARNEY, JM
通讯作者：
CARNEY, JM
影响因子：
4.8
作者：
Baker, MA;Lane, DJR;Lawen, A
通讯作者：
Lawen, A
DOI：
10.1016/j.nano.2010.01.010
发表时间：
2010-10-01
影响因子：
5.4
作者：
Colon, Jimmie;Hsieh, Nelson;Baker, Cheryl H.
通讯作者：
Baker, Cheryl H.
影响因子：
4.5
作者：
Arnold, S;Feng, ZQ;Niki, K
通讯作者：
Niki, K