HFPO-DA exacerbates acute ischemic stroke outcomes in rats via the CSNK2A1/GSK3B/NF-κB signaling pathway

The incidence of acute ischemic stroke (AIS) has shown a persistent upward trend in recent years. External factors, including chemical and plastic exposures, have been shown to worsen the prognosis of cerebrovascular diseases, especially AIS. Hexafluoropropylene oxide dimer acid (HFPO-DA), a prevalent plastic additive, is globally distributed. However, its bioaccumulation potential and neurovascular toxicity remain poorly understood. Using a mixed subchronic-acute aggravation model in middle cerebral artery occlusion/reperfusion (MCAO/R) rats, we found that HFPO-DA exposure dose-dependently exacerbated neurological deficits, enlarged infarct volumes, and intensified reactive microgliosis. Computationally, we prioritized the CSNK2A1/GSK3B/NF-κB signaling axis as a key mechanistic candidate, with bindings predicted by molecular docking. In vitro cellular thermal shift assays (CETSA) coupled with targeted siRNA knockdown in BV2 cells supported a direct physical interaction between HFPO-DA and CSNK2A1. Moreover, in vivo CSNK2A1 inhibition with CX-4945 partially reversed the HFPO-DA-induced aggravation of ischemic injury. Collectively, our results suggest that HFPO-DA might be a potent environmental driver of post-ischemic neuroinflammation, likely acting in part via the CSNK2A1/GSK3B/NF-κB signaling pathway. These results provide novel insights into the specific neurotoxic mechanisms of HFPO-DA, highlighting its potential health risks in susceptible populations.