Early label-free analysis of mitochondrial redox states by Raman spectroscopy predicts septic outcomes.
Early label-free analysis of mitochondrial redox states by Raman spectroscopy predicts septic outcomes.
复制标题
通过拉曼光谱对线粒体氧化还原状态进行早期无标记分析可预测败血症的结果
DOI：
10.1016/j.jare.2020.06.027
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发表时间：
2021-03
影响因子：
10.7
通讯作者：
Wang Q
中科院分区：
文献类型：
--
作者：
Wu M;Pu K;Jiang T;Zhai Q;Ma Z;Ma H;Xu F;Zhang Z;Wang Q
首次将共振拉曼光谱应用于脓毒症小鼠线粒体氧化还原状态的活体检测。使用共振拉曼光谱监测线粒体氧化还原状态对脓毒症期间死亡率的预测准确性高于乳酸水平，并且可能是在早期时间点预测脓毒症结局的新诊断标志物。共振拉曼光谱可以检测脓毒症中的线粒体功能障碍，并提供可以成为预防治疗的特异性靶点的生物标志物。由于缺乏早期预测脓毒症结局的生物标志物，脓毒症的死亡率仍然高得令人无法接受。线粒体氧化还原状态在这种情况下起着关键作用，并在脓毒症发展的早期受到干扰。在这里，我们假设通过共振拉曼光谱（RRS）可视化线粒体氧化还原状态可以在早期时间点识别脓毒症的结果。通过盲肠结扎穿孔（CLP）诱导脓毒症。分别在基线和CLP后30 min、1 h、2 h、4 h和6 h进行RRS分析，鉴定线粒体氧化还原状态。血乳酸水平作为脓毒症的预测指标进行了评估。我们的研究是第一个揭示在体内检测的线粒体氧化还原状态的可能性，在败血症小鼠使用RRS。拉曼还原的线粒体部分的峰面积，线粒体氧化还原状态的指标，在CLP后2小时波动显着。这种波动早于乳酸水平的变化。此外，这种波动比脓毒症期间的乳酸水平具有更高的死亡率预后准确性，并且可以成为预测脓毒症结局的新诊断标志物，根据截止值1.059，其灵敏度为80%，特异性为90%。目的探索脓毒症早期线粒体氧化还原状态的有效指标，并利用无创、无标记的共振拉曼光谱（RRS）技术在体内准确预测脓毒症的转归。用RRS法分别检测了大鼠原代骨骼肌细胞和离体腓肠肌线粒体的氧化还原状态。通过盲肠结扎穿孔（CLP）诱导脓毒症。分别在基线和CLP后30 min、1 h、2 h、4 h和6 h进行RRS分析，鉴定线粒体氧化还原状态。血乳酸水平作为脓毒症的预测指标进行了评估。进一步评估线粒体氧化还原状态对死亡率、炎症和器官功能障碍的预测相关性。线粒体氧化还原状态清楚地认识到在离体腓肠肌以及纯化的线粒体和原代骨骼肌细胞通过使用RRS。拉曼还原的线粒体部分的峰面积，线粒体氧化还原状态的指标，在CLP后2小时波动显着。这种波动早于乳酸水平的变化。此外，这种波动比脓毒症期间的乳酸水平具有更高的死亡率预后准确性，并且可以成为预测脓毒症结局的新诊断标志物，根据截止值1.059，其灵敏度为80%，特异性为90%。这项研究表明，早在2小时使用RRS监测线粒体氧化还原状态可以指示脓毒症小鼠的结果。这些数据可能有助于开发一个非侵入性的临床设备线粒体氧化还原状态使用床边RRS。
Resonance Raman spectroscopy was applied to in vivo detection of the mitochondrial redox state in septic mice for the first time. Monitoring mitochondrial redox states using resonance Raman spectroscopy had higher prognostic accuracy for mortality than the lactate level during sepsis and could be a novel diagnostic marker for predicting septic outcomes at an early time point. Resonance Raman spectroscopy could detect mitochondrial dysfunction in sepsis and provide a biomarker that can be a specific target of adjunctive treatment. Sepsis remains an unacceptably high mortality due to the lack of biomarkers for predicting septic outcomes in the early period. Mitochondrial redox states play a pivotal role in this condition and are disturbed early in the development of sepsis. Here, we hypothesized that visualizing mitochondrial redox states via resonance Raman spectroscopy (RRS) could identify septic outcomes at an early time point. Sepsis was induced by cecal ligation and puncture (CLP). We applied RRS analysis at baseline and 30 min, 1 h, 2 h, 4 h, and 6 h after CLP, and the mitochondrial redox states were identified. The levels of blood lactate as a predictor in sepsis were assessed. Our study is the first to reveal the possibility of in vivo detection of the mitochondrial redox state by using RRS in septic mice. The peak area for the Raman reduced mitochondrial fraction, the indicator of mitochondrial redox states, fluctuated significantly at 2 h after CLP. This fluctuation occurred earlier than the change in lactate level. Moreover, this fluctuation had higher prognostic accuracy for mortality than the lactate level during sepsis and could be a novel diagnostic marker for predicting septic outcomes according to the cutoff value of 1.059, which had a sensitivity of 80% and a specificity of 90%. To explore an effective indicator concerning mitochondrial redox states in the early stage of sepsis and to predict septic outcomes accurately in vivo using non-invasive and label-free Resonance Raman spectroscopy (RRS) analysis. Mitochondria, primary skeletal muscle cells andex-vivo muscles harvested from gastrocnemius were detected mitochondrial redox states respectively by using RRS. Sepsis was induced by cecal ligation and puncture (CLP). We applied RRS analysis at baseline and 30 min, 1 h, 2 h, 4 h, and 6 h after CLP, and the mitochondrial redox states were identified. The levels of blood lactate as a predictor in sepsis were assessed. The predictive correlation of mitochondrial redox states on mortality, inflammation and organ dysfunction was further assessed. Mitochondrial redox states were clearly recognized in ex-vivo gastrocnemius muscles as well as purified mitochondria and primary skeletal muscle cells by using RRS. The peak area for the Raman reduced mitochondrial fraction, the indicator of mitochondrial redox states, fluctuated significantly at 2 h after CLP. This fluctuation occurred earlier than the change in lactate level. Moreover, this fluctuation had higher prognostic accuracy for mortality than the lactate level during sepsis and could be a novel diagnostic marker for predicting septic outcomes according to the cutoff value of 1.059, which had a sensitivity of 80% and a specificity of 90%. This study demonstrated that monitoring mitochondrial redox states using RRS as early as 2 h could indicate outcomes in septic mice. These data may contribute to developing a non-invasive clinical device concerning mitochondrial redox states by using bedside-RRS.
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影响因子：
120.7
作者：
Jones, Alan E.;Shapiro, Nathan I.;Trzeciak, Stephen;Arnold, Ryan C.;Claremont, Heather A.;Kline, Jeffrey A.
通讯作者：
Kline, Jeffrey A.
影响因子：
3.7
作者：
Brazhe NA;Treiman M;Faricelli B;Vestergaard JH;Sosnovtseva O
通讯作者：
Sosnovtseva O
影响因子：
3.5
作者：
ADAR, F;ERECINSKA, M
通讯作者：
ERECINSKA, M
影响因子：
9.8
作者：
Nagy, J. A.;DiDonato, C. J.;Sanchez, B.
通讯作者：
Sanchez, B.
影响因子：
17.1
作者：
Perry, Dorothy A.;Salvin, Joshua W.;Kheir, John N.
通讯作者：
Kheir, John N.