Synergetic effect of mild hypothermia and antioxidant treatment on ROS-mediated neuron injury under oxygen-glucose deprivation investigated by scanning electrochemical microscopy.

Abstract

Ischemic stroke and reperfusion injury result in neuronal damage and dysfunction associated with oxidative stress, leading to overproduction of cellular reactive oxygen species (ROS) and reactive nitrogen species (RNS). In situ monitoring of the transient ROS and RNS effluxes during rapid pathologic processes is crucial for understanding the relationship between progression of cell damage and role of oxidative stress, and developing the corresponding neuroprotective strategies. Herein, we built oxygen glucose deprivation (OGD) and mild hypothermic (MH) models to mimic the in vitro conditions of ischemic stroke and MH treatment. We used scanning electrochemical microscopy (SECM) to in situ monitor H2O2 and nitric oxide (NO) effluxes from HT22 cells under the OGD and MH treatment conditions. Through quantitative analysis of the H2O2 and NO efflux results, we found that the cellular oxidative stress was primarily manifested through ROS release under OGD conditions, and the MH treatment partially suppressed the excessive H2O2 and NO production induced by reoxygenation. Moreover, the synergistic therapeutic effect of MH with antioxidant treatment significantly reduced the oxidative stress and enhanced the cell survival. Our work reveals the crucial role of oxidative stress in OGD and reperfusion processes, and the effective improvement of cell viability via combination of MH with antioxidants, proposing promising therapeutic interventions for ischemic stroke and reperfusion injury.