Aloe-emodin triggers ROS and Ca 2+ production and decreases the levels of mitochondrial membrane potential of human brain capillary endothelial cells

Authors

  • Ivanka Dimova Department of Medical Genetics, Medical Faculty, Medical University of Sofia; address: 2 Zdrave Str., Sofia, Bulgaria, post code 1431.
  • Svetla Danova The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences; address: 26 Akad. G. Bontchev Str., Sofia, Bulgaria, post code 1113
  • Ekaterina Nikolova Department of Medical Genetics, Medical Faculty, Medical University of Sofia; address: 2 Zdrave Str., Sofia, Bulgaria, post code 1431.
  • Miglena Koprinarova The Roumen Tsanev Institute of Molecular Biology, 21 Akad. G. Bontchev Str., Bulgarian Academy of Sciences, Sofia, Bulgaria, post code 1113

Keywords:

aloe-emodin, human brain capillary endothelial cells, reactive oxygen species, mitochondria, intracellular Ca 2

Abstract

The aim of this work was to investigate the mechanisms of cytotoxicity of phyto-hydroxyanthraquinone aloe-emodin (AE) on human brain microvascular endothelial cell line hCMEC/D3 and to assess the cellular response in the early stage of treatment in order to extend the knowledge of AE’s anti-angiogenic properties. The immortalized human brain capillary endothelial cells hCMEC/D3 were treated with a series of AE concentrations (5 - 200 μM) for a period of 24 hours. The cell viability was determined by MTS assay. The cellular adenosine triphosphate (ATP) levels were evaluated by CellTiter-Glo® luminescent assay. The intracellular reactive oxygen species (ROS) were determined by 2’,7’-dichlorofluorescein (CM- H2DCFDA) fluorescence assay. The mitochondrial membrane potential (MMP) was assessed using tetramethylrhodamine methyl ester (TMRM) staining, while Fluo-4 was used to measure 2 the intracellular free Ca 2+ concentrations inside living cells analysed by High Content Analysis using the Arrayscan VTI 740. Twenty-four- hour treatment of hCMEC/D3 cells with AE, in concentrations between 50 and 200 µM, decreased the cell viability as well as the intracellular ATP levels in a dose- dependent manner. Increased ROS production and disruption of the mitochondrial membrane potential have also been detected. Notably, AE at a concentration greater than 5 µM dramatically increased intracellular calcium levels. Our results collectively indicate that AE inhibits proliferation of human brain microvascular cells via a mechanism involving ROS generation, disruption of Ca 2+ homeostasis and mitochondrial damage.

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Published

30-09-2017

How to Cite

1.
Ivanka Dimova, Svetla Danova, Ekaterina Nikolova, Miglena Koprinarova. Aloe-emodin triggers ROS and Ca 2+ production and decreases the levels of mitochondrial membrane potential of human brain capillary endothelial cells. ijp [Internet]. 2017 Sep. 30 [cited 2024 Jun. 21];9(3):511-7. Available from: https://ijp.arjournals.org/index.php/ijp/article/view/564

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Original Research Articles