Zhao Liandong, Zhao Mingzhu, Wang Jing, Zhang Shuqi, Kong Rong-Mei, Yang Jianxin, Zhao Yan, Kong Weiheng
Key Laboratory of Life-Organic Analysis of Shandong Province, School of Chemistry and Chemical Engineering, Qufu Normal University, Qufu, 273165, P.R. China.
The Affiliated Hospital of Putian University, Putian University, Putian, 351100, P.R. China.
Mikrochim Acta. 2025 Jul 31;192(8):543. doi: 10.1007/s00604-025-07364-x.
Muscle injuries present a significant challenge in sports and physical activity, necessitating advanced diagnostic tools. Current invasive methods, such as muscle biopsies, are limited by procedural risks and lack of dynamic data. In this study, we developed a dual-channel fluorescent biosensor based on MoS nanosheets and fluorescence resonance energy transfer (FRET) for the simultaneous detection of miR-133a and ATP in living mouse myoblast cells. The design leverages the pathophysiological roles of miR-133a (a biomarker upregulated during muscle injury) and ATP (a critical indicator of energy metabolism). The biosensor incorporates FAM-labeled miR-133a-specific ssDNA and Cy5-labeled ATP aptamer probes immobilized on MoS nanosheets, achieving fluorescence quenching via FRET. Target binding triggered fluorescence recovery, enabling sensitive detection with limits of detection (LOD) of 3.3 pM for miR-133a and 0.33 µM for ATP. In vitro experiments confirmed high specificity and biocompatibility in mouse myoblasts, validated through confocal microscopy and flow cytometry. This platform provides a foundational tool for monitoring molecular dynamics associated with muscle injury, focusing on miR-133a and ATP levels in cellular environments. This work addresses limitations of traditional invasive diagnostics and lays the groundwork for potential applications in sports medicine research.
肌肉损伤在体育和身体活动中构成了重大挑战，这就需要先进的诊断工具。当前的侵入性方法，如肌肉活检，受到操作风险和缺乏动态数据的限制。在本研究中，我们基于二硫化钼纳米片和荧光共振能量转移（FRET）开发了一种双通道荧光生物传感器，用于在活的小鼠成肌细胞中同时检测miR-133a和三磷酸腺苷（ATP）。该设计利用了miR-133a（肌肉损伤期间上调的生物标志物）和ATP（能量代谢的关键指标）的病理生理作用。该生物传感器包含固定在二硫化钼纳米片上的羧基荧光素（FAM）标记的miR-133a特异性单链DNA和Cy5标记的ATP适配体探针，通过FRET实现荧光猝灭。目标物结合引发荧光恢复，能够对miR-133a进行灵敏检测，检测限（LOD）为3.3皮摩尔，对ATP的检测限为0.33微摩尔。体外实验证实了该生物传感器在小鼠成肌细胞中具有高特异性和生物相容性，并通过共聚焦显微镜和流式细胞术进行了验证。该平台为监测与肌肉损伤相关的分子动力学提供了一个基础工具，重点关注细胞环境中的miR-133a和ATP水平。这项工作解决了传统侵入性诊断的局限性，并为运动医学研究中的潜在应用奠定了基础。
