1. Academic Validation
  2. Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients

Spectrally and Time-Resolved Fluorescence Imaging of 22-NBD-Cholesterol in Human Peripheral Blood Mononuclear Cells in Chronic Kidney Disease Patients

  • Molecules. 2021 Nov 11;26(22):6800. doi: 10.3390/molecules26226800.
Ingrid Lajdova 1 Livia Ovsonkova 2 3 Viera Spustova 1 Adrian Oksa 1 Dusan Chorvat 2 Anton Mateasik 2 Alzbeta Marcek Chorvatova 2 3
Affiliations

Affiliations

  • 1 Department of Clinical and Experimental Pharmacology, Faculty of Medicine, Slovak Medical University, 83303 Bratislava, Slovakia.
  • 2 Department of Biophotonics, International Laser Centre of the Slovak Centre of Scientific and Technical Information, 84104 Bratislava, Slovakia.
  • 3 Department of Biophysics, Faculty of Natural Sciences, University of Ss. Cyril and Methodius, 91701 Trnava, Slovakia.
Abstract

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.

Keywords

22-NBD-cholesterol; CKD—chronic kidney disease; FLIM—fluorescence lifetime imaging microscopy; PBMC—peripheral blood mononuclear cells; spectrally resolved confocal microscopy; statins.

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