Department of Biomedical Engineering, School of Engineering, China Pharmaceutical University, 639 Longmian Road, Jiangning District, Nanjing, 211198, China.
Department of Pharmacy, The Affiliated Taizhou Second People's Hospital of Yangzhou University, No.27 Jiankang Road, Jiangyan District, Taizhou, 225500, China.
Biosens Bioelectron. 2025 Jan 1;267:116774. doi: 10.1016/j.bios.2024.116774. Epub 2024 Sep 11.
Abnormal lipid metabolism in glial cells is a key pathological feature of epilepsy. The identification of lipid droplets (LDs) is essential for investigating lipid metabolism, disease progression, and potential therapeutic interventions. Two-photon imaging technology enables real-time visualization of the spatial distribution and temporal dynamics of LDs in epilepsy models. In this study, we developed a novel two-photon excited dual-responsive near-infrared fluorescent probe, CabA, based on viscosity and polarity, to monitor dynamic changes in LDs. The fluorescence of CabA at 670 nm exhibits a significant increase in response to low polarity and high viscosity due to the twisted intramolecular charge transfer and intramolecular charge transfer mechanisms. The LDs-targeting capability of CabA at the cellular level and the process of LDs generation between neurons and astrocytes during the pathological advancement of epilepsy have been validated. In situ synchronous imaging experiments in epileptic and normal mice using CabA revealed abnormal LDs accumulation in the brain during seizures. Two-photon fluorescence imaging further demonstrated LDs accumulation in the brains of epileptic mice at a penetration depth of 100 μm. This study offers a valuable tool for enhancing the understanding of LDs in physiological and pathological processes, potentially aiding in the early diagnosis of epilepsy.
神经胶质细胞中的脂质代谢异常是癫痫的一个关键病理特征。脂滴（LDs）的鉴定对于研究脂质代谢、疾病进展和潜在的治疗干预措施至关重要。双光子成像技术能够实时可视化癫痫模型中 LDs 的空间分布和时间动态。在这项研究中，我们开发了一种基于粘度和极性的新型双光子激发双响应近红外荧光探针 CabA，用于监测 LDs 的动态变化。CabA 在 670nm 处的荧光由于扭曲的分子内电荷转移和分子内电荷转移机制，对低极性和高粘度表现出显著的增加。CabA 在细胞水平上对 LDs 的靶向能力以及在癫痫病理进展过程中神经元和星形胶质细胞之间 LDs 生成的过程已经得到了验证。使用 CabA 在癫痫和正常小鼠中进行的原位同步成像实验揭示了癫痫发作期间大脑中异常的 LDs 积累。双光子荧光成像进一步证明了癫痫小鼠大脑中 LDs 的积累，穿透深度为 100μm。这项研究提供了一种有价值的工具，用于增强对生理和病理过程中 LDs 的理解，可能有助于癫痫的早期诊断。
