Non-thermal plasma-induced DMPO-OH yields hydrogen peroxide
Non-thermal plasma-induced DMPO-OH yields hydrogen peroxide
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DOI：
10.1016/j.abb.2021.108901
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发表时间：
2021-05-11
影响因子：
3.9
通讯作者：
Toyokuni, Shinya
中科院分区：
文献类型：
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作者：
Okazaki, Yasumasa;Tanaka, Hiromasa;Toyokuni, Shinya
电子学的最新发展使非热等离子体（NTP）在医学上得到应用，它会产生活性氧（ROS）和活性氮（RNS），例如羟基自由基（（OH）-O-中心点）、过氧化氢（H2O2）、单线态氧（O-1（2））、超氧化物（O-2（中心点-））、 臭氧和一氧化氮在接近生理温度下。在临床前研究或人体临床试验中，NTP 可促进血液凝固，根除细菌、病毒和生物膜相关感染、伤口愈合和癌细胞死亡。为了阐明 NTP 在生物相容性还原剂存在下的溶液相生物效应，我们采用电子顺磁共振 (EPR) 光谱，使用自旋捕获探针 5,5-二甲基-1-吡咯啉-N-氧化物 (DMPO) 来量化 (OH)-O-中心点； O-1(2) 使用荧光探针；以及 O-2（中心点-）和 H2O2，在硫醇或 tempol 存在的情况下使用发光探针。 NTP 诱导的 (OH)-O-中心点被 2 或 5 mM DMPO 中的二硫苏糖醇 (DTT)、还原型谷胱甘肽 (GSH) 和氧化型谷胱甘肽 (GSSG) 显着清除。 NTP诱导的O-2(中心点-)被10μM DTT和GSH显着清除，而O-1(2)未被这些化合物有效清除。 GSSG 比 GSH 和 DTT 更有效地降解 H2O2，表明二硫键与 H2O2 发生反应。在 1-50 mM DMPO 存在下，NTP 诱导的 H2O2 量没有变化。在 1 和 10 mM DMPO 中观察到 tempol 浓度（50 和 100 μM）对 H2O2 产生的抑制作用，而在 50 mM DMPO 中则无效。此外，DMPO-OH 不与 tempol 相互作用。这些结果表明 DMPO 和 tempol 与 O-2（中心点-）竞争性反应。需要进一步的研究来阐明 NTP 诱导的 ROS 与生物分子之间的相互作用。
Recent developments in electronics have enabled the medical applications of non-thermal plasma (NTP), which elicits reactive oxygen species (ROS) and reactive nitrogen species (RNS), such as hydroxyl radical ((OH)-O-center dot), hydrogen peroxide (H2O2), singlet oxygen (O-1(2)), superoxide (O-2(center dot-)), ozone, and nitric oxide at near-physiological temperatures. In preclinical studies or human clinical trials, NTP promotes blood coagulation, eradication of bacterial, viral and biofilm-related infections, wound healing, and cancer cell death. To elucidate the solutionphase biological effects of NTP in the presence of biocompatible reducing agents, we employed electron paramagnetic resonance (EPR) spectroscopy to quantify (OH)-O-center dot using a spin-trapping probe, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO); O-1(2) using a fluorescent probe; and O-2(center dot-) and H2O2 using luminescent probes in the presence of thiols or tempol. NTP-induced (OH)-O-center dot was significantly scavenged by dithiothreitol (DTT), reduced glutathione (GSH), and oxidized glutathione (GSSG) in 2 or 5 mM DMPO. NTP-induced O-2(center dot-) was significantly scavenged by 10 mu M DTT and GSH, while O-1(2) was not efficiently scavenged by these compounds. GSSG degraded H2O2 more effectively than GSH and DTT, suggesting that the disulfide bonds reacted with H2O2. In the presence of 1-50 mM DMPO, NTP-induced H2O2 quantities were unchanged. The inhibitory effect of tempol concentration (50 and 100 mu M) on H2O2 production was observed in 1 and 10 mM DMPO, whereas it became ineffective in 50 mM DMPO. Furthermore, DMPO-OH did not interact with tempol. These results suggest that DMPO and tempol react competitively with O-2(center dot-). Further studies are warranted to elucidate the interaction between NTP-induced ROS and biomolecules.