State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, No.163 Xianlin Road, Nanjing, 210023, China.
Pancreas Center, The First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, 210029, China; Pancreas Institute of Nanjing Medical University, Nanjing, 210029, China.
Redox Biol. 2021 Oct;46:102068. doi: 10.1016/j.redox.2021.102068. Epub 2021 Jul 10.
Realizing the early diagnosis of acute liver injury has become a hotspot in recent years. Since the current indexes are not specific, developing novel probes with new recognition group remains a necessity. In this work, we developed a novel fluorescent probe, NNP, for the detection of ONOO in acute liver injury (ALI). The in vitro evaluation of NNP indicated the advantages including high sensitivity (LOD = 0.13 μM), rapid response (<25 s), naked-eye (red to yellow), and stability under various pH (4-10) and time (>48 h) conditions. Accordingly, NNP achieved the dose-dependent detection of ONOO in human hepatic stellate LX-2 cells. Further, in the ALI model mice, NNP could monitor the ONOO level in the occurrence of ALI in situ (in the liver region) with a steady performance. Subsequent immunohistochemical and imaging studies confirmed that NNP could achieve the detection of endogenous ONOO in ALI liver tissues. This work introduced a practical implement for the ONOO detection in ALI as well as a referable example for the establishment of molecular indexes in pre-clinical diagnosis.
实现急性肝损伤的早期诊断已成为近年来的研究热点。由于目前的指标特异性不强，因此开发具有新识别基团的新型探针仍然是必要的。在这项工作中，我们开发了一种用于检测急性肝损伤（ALI）中过氧亚硝酸盐（ONOO-）的新型荧光探针 NNP。NNP 的体外评价表明，它具有高灵敏度（LOD=0.13 μM）、快速响应（<25s）、裸眼（红色变为黄色）以及在各种 pH 值（4-10）和时间（>48h）条件下的稳定性等优点。因此，NNP 可以在人肝星状细胞 LX-2 细胞中实现对 ONOO-的剂量依赖性检测。此外，在 ALI 模型小鼠中，NNP 可以在原位（肝脏区域）监测 ALI 发生时的 ONOO-水平，性能稳定。随后的免疫组织化学和成像研究证实，NNP 可以检测到 ALI 肝组织中的内源性 ONOO-。这项工作为 ALI 中的 ONOO-检测引入了一种实用的方法，也为临床前诊断中分子指标的建立提供了一个可借鉴的范例。
