Bergapten attenuates oxidative stress and neuroinflammation to promote the survival and neuronal differentiation of neural stem/progenitor cells after ischemic insult.

Ischemic stroke remains a leading cause of death and disability worldwide. A significant challenge in recovery is the hostile microenvironment in the ischemic penumbra, characterized by excessive oxidative stress and neuroinflammation, which leads to massive neuronal death and impedes the regenerative potential of endogenous neural stem/progenitor cells (NSPCs). Bergapten, a natural coumarin derivative, has demonstrated antioxidant and anti-inflammatory properties in various disease models, suggesting its potential as a neuroprotective agent. This study aimed to investigate whether bergapten could protect against oxidative stress and inflammation in a simulated ischemic stroke model, and subsequently promote the survival, proliferation, and neuronal differentiation of NSPCs. We established in vitro models of oxidative stress using hydrogen peroxide (H2O2) treatment on primary mouse NSPCs and BV2 microglial cells. NSPCs were identified via immunofluorescence staining for Nestin, SRY-box transcription factor 2 (SOX2), and paired box protein 6 (PAX6). The effects of bergapten were evaluated by measuring: 1) intracellular reactive oxygen species (ROS) levels and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis in NSPCs; 2) intracellular ROS levels and M1/M2 polarization (inducible nitric oxide synthase (iNOS)/Arginase-1 (Arg-1)) in BV2 cells; and 3) neuronal or astroglial differentiation of NSPCs (neuronal class III β-tubulin (TUJ-1)) under stress conditions. H2O2 treatment induced significant oxidative stress and apoptosis in NSPCs, as evidenced by increased ROS levels and TUNEL-positive cells. It also triggered ROS production and a pro-inflammatory M1 phenotype (iNOS+) in BV2 microglia. Crucially, under H2O2-induced stress, NSPCs showed impaired differentiation into neurons (TUJ-1+ cells). Bergapten treatment effectively mitigated these detrimental effects: it reduced ROS levels and apoptosis in NSPCs, suppressed M1 polarization in BV2 cells, and, most importantly, rescued the capacity of NSPCs to differentiate into neurons. Our findings demonstrate that bergapten possesses potent antioxidant and anti-inflammatory properties in neural cell models of ischemic injury. By ameliorating the hostile microenvironment, bergapten enhances NSPC survival and facilitates their differentiation into neurons. This study provides the first experimental evidence supporting bergapten as a promising therapeutic candidate to promote endogenous neurogenesis and functional repair after ischemic stroke.