Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection.
Bortezomib is an effective enhancer for chemical probe-dependent superoxide detection.
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DOI：
10.3389/fmed.2022.941180
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发表时间：
2022
影响因子：
3.9
通讯作者：
中科院分区：
文献类型：
--
作者：
已经开发出用于检测活性氧的各种化学探针来检查与不同病理相关的氧化应激。 L-012是一种基于鲁米诺的化学发光探针，因其高灵敏度而被广泛用于检测细胞外超氧化物。我们在此证明，肽硼酸蛋白酶体抑制剂硼替佐米与L-012的共同应用显着增加了其发光而不影响背景。在表达 NADPH 氧化酶的细胞和依赖黄嘌呤氧化酶的无细胞超氧化物生成系统中，检测到 L-012 的总发光增加了 5 倍以上，但在它们的背景中却没有增加。因此，硼替佐米提高了信号背景比并改善了低水平超氧化物的检测。另一种肽硼酸蛋白酶体抑制剂MLN2238的应用也增强了L-012的发光。相比之下，环氧酮蛋白酶体抑制剂卡非佐米并没有增加发光，这表明硼替佐米的作用取决于肽硼酸的化学结构，而不是其药理作用。硼替佐米诱导的增强似乎对超氧化物的检测具有特异性，因为 Amplex Red/HRP 对 H2O2 的检测不受硼替佐米应用的影响。在超氧化物特异性氧化产物2-羟基乙锭（2-OH-E+）的定量检测中，硼替佐米的应用导致2-OH-E+水平增加2倍。因此，硼替佐米使细胞和无细胞系统中超氧化物的检测变得敏感，突出了含有肽硼酸的化合物作为超氧化物检测的强大增强剂的新特征。
Various chemical probes for the detection of reactive oxygen species have been developed to examine oxidative stress associated with different pathologies. L-012, a luminol-based chemiluminescent probe, is widely used to detect extracellular superoxide because of its high sensitivity. We herein demonstrated that the co-application of the peptide boronic acid proteasome inhibitor, bortezomib, with L-012 significantly increased its luminescence without affecting the background. More than a 5-fold increase was detected in the total luminescence of L-012 in both NADPH oxidase-expressing cells and the xanthine oxidase-dependent cell-free superoxide generation system, but not in their background. Therefore, bortezomib increased the signal-to-background ratio and improved the detection of low levels of superoxide. The application of MLN2238, another peptide boronic acid proteasome inhibitor, also enhanced the luminescence of L-012. In contrast, carfilzomib, an epoxyketone proteasome inhibitor, did not increase luminescence, suggesting that the effects of bortezomib depend on the chemical structure of the peptide boronic acid, but not on its pharmacological effects. Bortezomib-induced enhancements appeared to be specific to the detection of superoxide because the detection of H2O2 by Amplex Red/HRP was not affected by the application of bortezomib. In the quantitative detection of the superoxide-specific oxidative product 2-hydroxyethidium (2-OH-E+), the application of bortezomib resulted in a 2-fold increase in the level of 2-OH-E+. Therefore, bortezomib sensitizes the detection of superoxide in both cell-based and cell-free systems, highlighting a novel feature of compounds containing the peptide boronic acid as powerful enhancers for the detection of superoxide.
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影响因子：
7.4
作者：
Zielonka, Jacek;Lambeth, J. David;Kalyanaraman, Balaraman
通讯作者：
Kalyanaraman, Balaraman
影响因子：
4.1
作者：
Arakawa, Noriaki;Katsuyama, Masato;Yabe-Nishimura, Chihiro
通讯作者：
Yabe-Nishimura, Chihiro
影响因子：
7.4
作者：
Daiber, A;August, M;Munzel, T
通讯作者：
Munzel, T
影响因子：
2.6
作者：
Zielonka J;Hardy M;Michalski R;Sikora A;Zielonka M;Cheng G;Ouari O;Podsiadły R;Kalyanaraman B
通讯作者：
Kalyanaraman B
DOI：
10.2147/dddt.s93602
发表时间：
2016
期刊：
Drug design, development and therapy
影响因子：
--
作者：
Muz B;Ghazarian RN;Ou M;Luderer MJ;Kusdono HD;Azab AK
通讯作者：
Azab AK