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

Autor: Ingrid Lajdova, Adrian Oksa, Anton Mateasik, Dusan Chorvat, Alzbeta Chorvatova, Viera Spustova, Livia Ovsonkova
Rok vydání: 2021
Předmět:
Pathology
medicine.medical_specialty
Fluorescence-lifetime imaging microscopy
FLIM—fluorescence lifetime imaging microscopy
Pharmaceutical Science
Organic chemistry
Pilot Projects
Peripheral blood mononuclear cell
Article
Analytical Chemistry
law.invention
statins
spectrally resolved confocal microscopy
QD241-441
Confocal microscopy
law
Drug Discovery
medicine
Humans
Physical and Theoretical Chemistry
Renal Insufficiency
Chronic
Fluorescent Dyes
CKD—chronic kidney disease
Microscopy
Confocal
Chemistry
Cell Membrane
Lipid metabolism
Intracellular Membranes
PBMC—peripheral blood mononuclear cells
medicine.disease
Lipid Metabolism
Fluorescence
Healthy Volunteers
Spectral imaging
4-Chloro-7-nitrobenzofurazan
Cholesterol
Chemistry (miscellaneous)
Leukocytes
Mononuclear
Molecular Medicine
lipids (amino acids
peptides
and proteins)
Time-resolved spectroscopy
Hydroxymethylglutaryl-CoA Reductase Inhibitors
22-NBD-cholesterol
Kidney disease
Zdroj: Molecules
Volume 26
Issue 22
Molecules, Vol 26, Iss 6800, p 6800 (2021)
ISSN: 1420-3049
Popis: 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.
Databáze: OpenAIRE
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