Cargando…

Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium

Chloroquine (CQ) is an antimalarial drug known to inhibit autophagy flux by impairing autophagosome–lysosome fusion. We hypothesized that autophagy flux altered by CQ has a considerable influence on the lipid composition of endothelial cells. Thus, we investigated endothelial responses induced by CQ...

Descripción completa

Detalles Bibliográficos
Autores principales: Bik, Ewelina, Mateuszuk, Lukasz, Orleanska, Jagoda, Baranska, Malgorzata, Chlopicki, Stefan, Majzner, Katarzyna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7957661/
https://www.ncbi.nlm.nih.gov/pubmed/33673688
http://dx.doi.org/10.3390/ijms22052401
_version_ 1783664700256419840
author Bik, Ewelina
Mateuszuk, Lukasz
Orleanska, Jagoda
Baranska, Malgorzata
Chlopicki, Stefan
Majzner, Katarzyna
author_facet Bik, Ewelina
Mateuszuk, Lukasz
Orleanska, Jagoda
Baranska, Malgorzata
Chlopicki, Stefan
Majzner, Katarzyna
author_sort Bik, Ewelina
collection PubMed
description Chloroquine (CQ) is an antimalarial drug known to inhibit autophagy flux by impairing autophagosome–lysosome fusion. We hypothesized that autophagy flux altered by CQ has a considerable influence on the lipid composition of endothelial cells. Thus, we investigated endothelial responses induced by CQ on human microvascular endothelial cells (HMEC-1). HMEC-1 cells after CQ exposure were measured using a combined methodology based on label-free Raman and fluorescence imaging. Raman spectroscopy was applied to characterize subtle chemical changes in lipid contents and their distribution in the cells, while the fluorescence staining (LipidTox, LysoTracker and LC3) was used as a reference method. The results showed that CQ was not toxic to endothelial cells and did not result in the endothelial inflammation at concentrations of 1–30 µM. Notwithstanding, it yielded an increased intensity of LipidTox, LysoTracker, and LC3 staining, suggesting changes in the content of neutral lipids, lysosomotropism, and autophagy inhibition, respectively. The CQ-induced endothelial response was associated with lipid accumulation and was characterized by Raman spectroscopy. CQ-induced autophagosome accumulation in the endothelium is featured by a pronounced alteration in the lipid profile, but not in the endothelial inflammation. Raman-based assessment of CQ-induced biochemical changes offers a better understanding of the autophagy mechanism in the endothelial cells.
format Online
Article
Text
id pubmed-7957661
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-79576612021-03-16 Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium Bik, Ewelina Mateuszuk, Lukasz Orleanska, Jagoda Baranska, Malgorzata Chlopicki, Stefan Majzner, Katarzyna Int J Mol Sci Article Chloroquine (CQ) is an antimalarial drug known to inhibit autophagy flux by impairing autophagosome–lysosome fusion. We hypothesized that autophagy flux altered by CQ has a considerable influence on the lipid composition of endothelial cells. Thus, we investigated endothelial responses induced by CQ on human microvascular endothelial cells (HMEC-1). HMEC-1 cells after CQ exposure were measured using a combined methodology based on label-free Raman and fluorescence imaging. Raman spectroscopy was applied to characterize subtle chemical changes in lipid contents and their distribution in the cells, while the fluorescence staining (LipidTox, LysoTracker and LC3) was used as a reference method. The results showed that CQ was not toxic to endothelial cells and did not result in the endothelial inflammation at concentrations of 1–30 µM. Notwithstanding, it yielded an increased intensity of LipidTox, LysoTracker, and LC3 staining, suggesting changes in the content of neutral lipids, lysosomotropism, and autophagy inhibition, respectively. The CQ-induced endothelial response was associated with lipid accumulation and was characterized by Raman spectroscopy. CQ-induced autophagosome accumulation in the endothelium is featured by a pronounced alteration in the lipid profile, but not in the endothelial inflammation. Raman-based assessment of CQ-induced biochemical changes offers a better understanding of the autophagy mechanism in the endothelial cells. MDPI 2021-02-27 /pmc/articles/PMC7957661/ /pubmed/33673688 http://dx.doi.org/10.3390/ijms22052401 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bik, Ewelina
Mateuszuk, Lukasz
Orleanska, Jagoda
Baranska, Malgorzata
Chlopicki, Stefan
Majzner, Katarzyna
Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium
title Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium
title_full Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium
title_fullStr Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium
title_full_unstemmed Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium
title_short Chloroquine-Induced Accumulation of Autophagosomes and Lipids in the Endothelium
title_sort chloroquine-induced accumulation of autophagosomes and lipids in the endothelium
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7957661/
https://www.ncbi.nlm.nih.gov/pubmed/33673688
http://dx.doi.org/10.3390/ijms22052401
work_keys_str_mv AT bikewelina chloroquineinducedaccumulationofautophagosomesandlipidsintheendothelium
AT mateuszuklukasz chloroquineinducedaccumulationofautophagosomesandlipidsintheendothelium
AT orleanskajagoda chloroquineinducedaccumulationofautophagosomesandlipidsintheendothelium
AT baranskamalgorzata chloroquineinducedaccumulationofautophagosomesandlipidsintheendothelium
AT chlopickistefan chloroquineinducedaccumulationofautophagosomesandlipidsintheendothelium
AT majznerkatarzyna chloroquineinducedaccumulationofautophagosomesandlipidsintheendothelium