A near-infrared fluorescent probe for dynamic HClO monitoring in epilepsy and high-throughput discovery of natural neuroprotectants

Epilepsy is a neurodegenerative disease closely associated with mitochondrial oxidative stress and neuronal damage, posing a grave threat to human health. Hypochlorous acid (HClO), a potent oxidant, has been demonstrated to play a critical role in the occurrence and progression of epilepsy. Accordingly, the development of a mitochondrial-targeted responsive HClO fluorescent probe could serve as a powerful tool for the early diagnosis of epilepsy and high-throughput screening (HTS) of natural neuroprotective. Herein, we report a mitochondria-targeted fluorescent probe (MB-Mito) with excellent selectivity, rapid activation kinetics, and blood-brain barrier (BBB) permeability, which is used for real-time and in situ tracking of HClO fluctuations in epilepsy models. More significantly, a visual HTS platform for natural neuroprotective agents was established based on MB-Mito, and an effective neuroprotective agent, myricetin, was identified from 47 natural compounds with potential antioxidant activity. Mechanistic studies have demonstrated that myricetin activates the ERK/Nrf2/HO-1 signaling pathway to mitigate mitochondrial oxidative stress, thereby providing precise treatment for epilepsy. All studies have confirmed that MB-Mito is a reliable tool for precise monitoring of HClO, drug HTS and mechanism research, promoting advances in the diagnosis and treatment of epilepsy.