Analysis of reactive oxygen and nitrogen species generated in three liquid media by low temperature helium plasma jet.
Analysis of reactive oxygen and nitrogen species generated in three liquid media by low temperature helium plasma jet.
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DOI：
10.1038/s41598-017-04650-4
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发表时间：
2017-07-04
影响因子：
4.6
通讯作者：
Merbahi N
中科院分区：
文献类型：
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作者：
Chauvin J;Judée F;Yousfi M;Vicendo P;Merbahi N
为了鉴别等离子体活化培养基（PAM）中形成的水相物质，对活性氧和氮物质（ROS、RNS）进行了定量研究，并与Milli-Q水和不含和含胎牛血清的培养基进行了比较。电子顺磁共振，荧光和比色分析，用于确定和定量的氦等离子体射流在这些液体中产生的自由基。结果清楚地表明，形成的活性氧，如羟基自由基，超氧阴离子自由基和单线态氧的微摩尔浓度范围内的顺序。一氧化氮，过氧化氢和亚硝酸根-硝酸根阴离子（在几百微摩尔的范围内）是在PAM中观察到的主要物种。介质的组成对等离子体处理过程中溶液的pH值、所产生的不同RONS的稳定性以及它们与生物分子的反应性具有重大影响。为了强调血浆与复杂介质的相互作用，还使用蛋氨酸、酪氨酸、色氨酸和精氨酸研究了通过质谱法的氨基酸降解。所有这些成分，例如长寿命的RONS和氧化的生物化合物，可能有助于PAM的细胞毒性作用。这项研究提供了机制的见解与PAM处理后的细胞死亡的机制。
In order to identify aqueous species formed in Plasma activated media (PAM), quantitative investigations of reactive oxygen and nitrogen species (ROS, RNS) were performed and compared to Milli-Q water and culture media without and with Fetal Calf Serum. Electron paramagnetic resonance, fluorometric and colorimetric analysis were used to identify and quantify free radicals generated by helium plasma jet in these liquids. Results clearly show the formation of ROS such as hydroxyl radical, superoxide anion radical and singlet oxygen in order of the micromolar range of concentrations. Nitric oxide, hydrogen peroxide and nitrite-nitrate anions (in range of several hundred micromolars) are the major species observed in PAM. The composition of the medium has a major impact on the pH of the solution during plasma treatment, on the stability of the different RONS that are produced and on their reactivity with biomolecules. To emphasize the interactions of plasma with a complex medium, amino acid degradation by means of mass spectrometry was also investigated using methionine, tyrosine, tryptophan and arginine. All of these components such as long lifetime RONS and oxidized biological compounds may contribute to the cytotoxic effect of PAM. This study provides mechanistic insights into the mechanisms involved in cell death after treatment with PAM.