Kim Su Jung, Park Sun Young, Yoon Shin A, Kim Changshin, Kang Chulhun, Lee Min Hee
Department of Chemistry, Sookmyung Women's University, Seoul 04310, Korea.
The School of East-West Medical Science, Kyung Hee University, Yongin 17104, Korea.
Anal Chem. 2021 Mar 16;93(10):4391-4397. doi: 10.1021/acs.analchem.0c04019. Epub 2021 Feb 22.
Intracellular viscosity is a physicochemical factor that determines the outcomes of various biological processes, while nitric oxide (NO) is an essential signaling molecule that controls many cellular processes, including oxidative stress. Anticipating that both may be interrelated with a variety of pathologies, their simultaneous measurement would be highly valuable for the investigation of the pathological condition of cells. However, the development of a sensor for such simultaneous detection has not been attempted yet. Herein, we present the synthesis of naphthalimide-4-(4-nitrophenyl)thiosemicarbazide, probe , and its application to living cells under conditions of lipopolysaccharide or nystatin treatment, adopted as oxidative stress and altered intracellular viscosity models, respectively. The probe showed increased fluorescence in response to elevation of viscosity and NO levels at 470 and 550 nm, respectively, in the solution studies. When the probe was used for a confocal microscopic study of HeLa cells under stressed conditions, simultaneous monitoring of viscosity and NO level elevations was possible through fluorescence imaging using band-pass filters of 420-475 and 505-600 nm, respectively, upon excitation at a wavelength of 405 nm. Interestingly, both the cellular viscosity and NO levels increased together under lipopolysaccharide or nystatin treatment. Therefore, we suggest that probe is a fluorescent chemical probe that enables the monitoring of alterations in intracellular viscosity and NO levels in living cells, which would be valuable in studies of various cellular damage models.
细胞内粘度是决定各种生物过程结果的物理化学因素，而一氧化氮（NO）是控制许多细胞过程（包括氧化应激）的重要信号分子。预计两者可能与多种病理状况相关，同时测量它们对于研究细胞的病理状况将具有很高的价值。然而，尚未尝试开发用于这种同时检测的传感器。在此，我们展示了萘二甲酰亚胺-4-（4-硝基苯基）硫代氨基脲探针的合成及其在脂多糖或制霉菌素处理条件下对活细胞的应用，分别将其用作氧化应激和细胞内粘度改变的模型。在溶液研究中，该探针分别在470和550nm处对粘度和NO水平的升高表现出荧光增强。当该探针用于应激条件下HeLa细胞的共聚焦显微镜研究时，在405nm波长激发下，分别使用420 - 475和505 - 600nm的带通滤光片通过荧光成像可以同时监测粘度和NO水平的升高。有趣的是，在脂多糖或制霉菌素处理下，细胞粘度和NO水平同时升高。因此，我们认为探针是一种荧光化学探针，能够监测活细胞内细胞粘度和NO水平的变化，这在各种细胞损伤模型的研究中具有重要价值。
