Department of Clinical and Experimental Pharmacology, Faculty of Medicine, Slovak Medical University, 83303 Bratislava, Slovakia.
Department of Biophotonics, International Laser Centre of the Slovak Centre of Scientific and Technical Information, 84104 Bratislava, Slovakia.
Molecules. 2021 Nov 11;26(22):6800. doi: 10.3390/molecules26226800.
The interaction of the fluorescent probe 22-NBD-cholesterol with membranes of human peripheral blood mononuclear cells (PBMC) was tested by time- and spectrally resolved fluorescence imaging to monitor the disturbance of lipid metabolism in chronic kidney disease (CKD) and its treatment with statins. Blood samples from healthy volunteers (HV) and CKD patients, either treated or untreated with statins, were compared. Spectral imaging was done using confocal microscopy at 16 spectral channels in response to 458 nm excitation. Time-resolved imaging was achieved by time-correlated single photon counting (TCSPC) following excitation at 475 nm. The fluorescence of 22-NBD-cholesterol was mostly integrated into plasmatic membrane and/or intracellular membrane but was missing from the nuclear region. The presence of two distinct spectral forms of 22-NBD-cholesterol was uncovered, with significant variations between studied groups. In addition, two fluorescence lifetime components were unmasked, changing in CKD patients treated with statins. The gathered results indicate that 22-NBD-cholesterol may serve as a tool to study changes in the lipid metabolism of patients with CKD to monitor the effect of statin treatment.
采用时间分辨和光谱分辨荧光成像技术研究荧光探针 22-NBD-胆固醇与人类外周血单个核细胞（PBMC）膜的相互作用，以监测慢性肾脏病（CKD）及其他汀类药物治疗过程中脂质代谢的紊乱。比较了来自健康志愿者（HV）和 CKD 患者的血液样本，这些患者要么接受他汀类药物治疗，要么未接受他汀类药物治疗。采用共聚焦显微镜在 16 个光谱通道中进行光谱成像，以响应 458nm 的激发。通过在 475nm 激发后进行时间相关单光子计数（TCSPC）实现时间分辨成像。22-NBD-胆固醇的荧光主要整合到质膜和/或细胞内膜中，但不存在于核区域。发现 22-NBD-胆固醇存在两种不同的光谱形式，在研究组之间存在显著差异。此外，还揭示了两种荧光寿命成分的变化，这些变化发生在接受他汀类药物治疗的 CKD 患者中。收集到的结果表明，22-NBD-胆固醇可用作研究 CKD 患者脂质代谢变化的工具，以监测他汀类药物治疗的效果。
