State Key Laboratory of Materials-Oriented Chemical Engineering, College of Chemical Engineering, Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing Tech University, Nanjing 211816, China.
Institute of Translational Medicine, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
Anal Methods. 2024 Jun 27;16(25):4054-4059. doi: 10.1039/d4ay00662c.
Liver diseases are a growing public health concern and the development of non-alcoholic fatty liver disease (NAFLD) has a significant impact on human metabolism. Butyrylcholinesterase (BChE) is a vital biomarker for NAFLD, making it crucial to monitor BChE activity with high sensitivity and selectivity. In this study, we designed and synthesized a range of benzorhodol-derived far-red/near-infrared fluorescent probes, FRBN-B, NF-SB, and NF-B, for the quantitative detection and imaging of BChE. These probes differed in the size of their conjugated systems and in the number of incorporated cyclopropanecarboxylates, acting as the recognition site for BChE. Comprehensive characterization showed that FRBN-B and NF-SB fluorescence was triggered by BChE-mediated hydrolysis, while an additional cyclopropanecarboxylate in NF-B impeded the fluorescence release. High selectivity towards BChE was observed for FRBN-B and NF-SB, with a detection limit of 7.2 × 10 U mL for FRBN-B and 1.9 × 10 U mL for NF-SB. The probes were further employed in the evaluation of BChE inhibitor efficacy and imaging of intracellular BChE activity. Additionally, FRBN-B was utilized for imaging the BChE activity level in liver tissues in zebrafish, demonstrating its potential as a diagnostic tool for NAFLD.
肝脏疾病是一个日益严重的公共卫生问题，非酒精性脂肪性肝病 (NAFLD) 的发展对人类代谢有重大影响。丁酰胆碱酯酶 (BChE) 是 NAFLD 的一个重要生物标志物，因此，需要用高灵敏度和高选择性来监测 BChE 的活性。在这项研究中，我们设计并合成了一系列苯并罗丹衍生的远红/近红外荧光探针，FRBN-B、NF-SB 和 NF-B，用于 BChE 的定量检测和成像。这些探针的共轭体系大小和整合的环丙烷羧酸数目不同，作为 BChE 的识别位点。综合表征表明，FRBN-B 和 NF-SB 的荧光是由 BChE 介导的水解触发的，而 NF-B 中的另一个环丙烷羧酸则阻碍了荧光的释放。FRBN-B 和 NF-SB 对 BChE 具有高选择性，FRBN-B 的检测限为 7.2×10 U mL，NF-SB 的检测限为 1.9×10 U mL。这些探针进一步用于评估 BChE 抑制剂的功效和细胞内 BChE 活性的成像。此外，FRBN-B 还用于在斑马鱼的肝组织中成像 BChE 的活性水平，表明其作为 NAFLD 诊断工具的潜力。
