Reactive Oxygen Species: A Key Hallmark of Cardiovascular Disease
Table 1
Direct methods for detection of ROS in CVDs.
Methods
ROS detected
Applications/mechanism
Reference
Fluorescent protein-based redox probes
Cytoplasmic and mitochondrial H2O2
Used to detect redox status and ROS by introducing adenoviruses or plasmids inside cells. Afterwards, cells form chimeric proteins efficient to detect alteration in the redox status or ROS.
Dihydroethidium (DHE) and mitochondrion-targeted probe mitoSOX
Cellular and mitochondrial
Can detect mitochondrial by adding a triphenylphosphonium group for promoting its collection in the mitochondria. Similar to DHE, mitoSOX reacts with to give 2-hydroxy-mito-ethidium (2-OH-Mito-E+) so as to be identified and measured using HPLC.
As probes have a fluorophore which is secured by boronate, when subjected to H2O2, the boronate encounters a nucleophilic attack, followed by its displacement from the fluorophore, thus causing emission of light.
Detection is done in vivo by infusion of cyclic hydroxylamines or nitrone spin traps, followed by ex vivo study of the tissue or blood using X-band (9 GHz) electron spin resonance spectroscopy.