Epileptic brain fluorescent imaging reveals apigenin can relieve the myeloperoxidase-mediated oxidative stress and inhibit ferroptosis

Abstract

Myeloperoxidase (MPO)-mediated oxidative stress has been suggested to play an important role in the pathological dysfunction of epileptic brains. However, there is currently no robust brain-imaging tool to detect real-time endogenous hypochlorite (HClO) generation by MPO or a fluorescent probe for rapid high-throughput screening of antiepileptic agents that control the MPO-mediated chlorination stress. Herein, we report an efficient two-photon fluorescence probe (named HCP) for the real-time detection of endogenous HClO signals generated by MPO in the brain of kainic acid (KA)-induced epileptic mice, where HClO-dependent chlorination of quinolone fluorophore gives the enhanced fluorescence response. With this probe, we visualized directly the endogenous HClO fluxes generated by the overexpression of MPO activity in vivo and ex vivo in mouse brains with epileptic behaviors. Notably, by using HCP, we have also constructed a high-throughput screening approach to rapidly screen the potential antiepileptic agents to control MPO-mediated oxidative stress. Moreover, from this screen, we identified that the flavonoid compound apigenin can relieve the MPO-mediated oxidative stress and inhibit the ferroptosis of neuronal cells. Overall, this work provides a versatile fluorescence tool for elucidating the role of HClO generation by MPO in the pathology of epileptic seizures and for rapidly discovering additional antiepileptic agents to prevent and treat epilepsy.

A comparative study of the phenotype with kainic acid-induced seizure in DBA/2 mice from three different sources

“…Diverse mechanisms have been uncovered with laboratory animal models, including those as a result of toxicity and drug efficacy studies. DBA/2 mice have been previously used in KA-induced seizure studies [ 7 – 9 ]. However, no comparative studies have been conducted regarding DBA/2 mice from separate sources.…”

“…Rodents, including DBA/2 mice, have generally been used as animal models for a long time. KA-induced seizure studies are performed using DBA/2 mice [ 6 – 9 ] and include toxicity, brain and biological response tests [ 10 – 14 ]. Among laboratory animals, DBA/2 mouse is a commonly used model.…”

Recurring Real-Time Monitoring of Inflammations in Living Mice with a Chemiluminescent Probe for Hypochlorous Acid

“…Collectively, the robust performance of cell imaging in 96-well plate could be further explored to develop high-throughput cellular assays for HOCl stimulating reagents or scavengers. 14 Moreover, as no laser is needed for chemiluminescent imaging, those autofluorescent drugs and compounds in high-throughput screening library could in principle be screened by using HOCl-CL-510 without worrying about the false positive hits due to auto-fluorescence.…”

Iron Metabolism and Ferroptosis in Epilepsy

“…Fisetin reduces lipid peroxides and retains Na (+), K (+)-ATPase activities in PTE, and executes an antiepileptic role in ironinduced epileptic rat models by inhibiting oxidative stress and reduces the cognitive dysfunction related to epileptic seizures (Jharana et al, 2017). Epileptic brain fluorescent imaging revealed that apigenin relieves myeloperoxidase-mediated oxidative stress and inhibits ferroptosis (Shao et al, 2020).…”