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Development of Novel Apparatus for Singlet Oxygen Detection and Its Application as Biological Tool

新型单线态氧检测装置的研制及其作为生物工具的应用

基本信息

批准号：
06557139
负责人：
NAGANO Tetsuo
金额：
$ 3.9万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

项目摘要

Singlet oxygen, a highy reacive and toxic intermediate, plays an important role in many photochemical and photobiological processes and has been postulated to be a major species mediating photochemical action in biological systems. Many attempts have been made to find evidence of singlet oxygen generation in various photosensitized reactions and enzymatic systems. Compared with the known nonspecific techniques for the detection of singlet oxygen, direct obervation of the emission at 1268 nm has proven to be the most reliable approach for singlet oxygen detection. We have constructed a sensitive near-infrared emission spectrometer with germanium detector, which scans from 1000 nm to 1600 nm. We applied this apparatus for detection of singlet oxygen produced by laser-excited photosensitive dyes. For example, the data about new quinolones will be shown below.New quinolones have potent anti-bacterial activity based on the inhibition of bacterial DNA gyrase and have been widely used as a cu … More rative medicine. However, some of them have been reported to induce photodermatosis as side effect. We have examined 1)the production of reactive oxygen species (ROS) mediated by new quinolones shown below and 2)participation of ROS in the phototoxicity. We employed the near-infrared (1268 nm) emission spectrometer to detect singlet oxygen specific emission, and NBT reduction method to detect superoxide. We compared ROS formation ability of the new quinolones quantitatively. Since it is known that new quinolones are able to interact with DNA,we also investigated the photodynamic DNA strand breaking activity that can estimate the phototoxicity of the new quinolones. All the new quinolones investigated in this study were found to produce singlet oxygen, and they also produced superoxide except SPFX.It has been little reported that colorless or pale color compounds act as photosensitizer that produces ROS.They also had photodynamic DNA strand breaking activity. The main active species was probably singlet oxygen, because NaN,inhibited the activity strongly. Less

单线态氧是一种高活性、高毒性的中间体，在许多光化学和光生物学过程中起着重要作用，被认为是生物系统中介导光化学作用的主要物种。已经进行了许多尝试来寻找在各种光敏化反应和酶系统中单线态氧产生的证据。与已知的非特异性单重态氧检测技术相比，直接观察1268 nm处的发射已被证明是最可靠的单重态氧检测方法。我们研制了一台灵敏的锗探测器近红外发射光谱仪，扫描范围为1000 ~ 1600 nm。我们将此装置应用于激光激发光敏染料产生的单线态氧的检测。例如，新喹诺酮类药物的数据如下所示，新喹诺酮类药物基于对细菌DNA旋转酶的抑制而具有强效抗菌活性，已被广泛用作抗菌药物。 ...更多信息治疗药物然而，其中一些已被报道诱发光照皮肤病作为副作用。我们已经检查了1）由下面所示的新喹诺酮介导的活性氧（ROS）的产生和2）ROS在光毒性中的参与。我们采用近红外（1268 nm）发射光谱仪检测单线态氧的比发射，NBT还原法检测超氧阴离子。我们定量比较了新喹诺酮类药物的活性氧形成能力。由于已知新喹诺酮类药物能够与DNA相互作用，我们还研究了光动力学DNA链断裂活性，可以估计新喹诺酮类药物的光毒性。本研究中所研究的新喹诺酮类化合物除SPFX外，均能产生单线态氧，也能产生超氧阴离子，无色或浅色化合物作为光敏剂产生ROS的报道较少，它们也具有光动力学DNA链断裂活性。主要活性物种可能是单线态氧，因为NaN强烈抑制活性。少