Low-intensity transcranial ultrasound (LITUS) exerts neuroprotective effects by modulating inflammatory responses via the Piezo1-Dkk3/Ca 2+ /PI3K-Akt signaling pathway

Abstract

Abstract Background Post-cardiac arrest brain injury (PCABI) contributes to unfavorable outcomes following cardiopulmonary resuscitation (CPR) significantly, with effective therapeutic interventions for PCABI remain elusive. Low-intensity transcranial ultrasound (LITUS) has exhibited neuroprotective effects in various disease models, yet the precise mechanisms underlying these effects have not been fully elucidated. Consequently, the therapeutic potential of ultrasound stimuli in PCABI and the underlying mechanisms necessitate further exploration. Methods In vivo, a murine model of cardiac arrest (CA) was established and subjected to LITUS. Neurological function was assessed through behavioral tests and neurofunctional scoring. Hematoxylin and Eosin (HE) staining was employed to evaluate brain injury, while proteomic analysis was utilized to identify potential target proteins and pathways. Cells involved in pharmacological interventions or gene knockdown were subjected to oxygen/glucose deprivation/reoxygenation to mimic ischemia-reperfusion conditions in the brain. Protein-protein interactions were confirmed through co-immunoprecipitation. Intracellular calcium (Ca2+) influx was quantified using live-cell calcium imaging and flow cytometry. Cell viability was assessed using the Cell Counting Kit-8 (CCK8) assay. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to evaluate the expression of targeted genes. The expression of related proteins was detected through immunofluorescence and Western blotting assays. Results LITUS administration significantly improves 24-hour survival rates, promotes neurological function recovery, attenuates brain injury, and activates neurons in mice subjected to CPR. In vitro experiments revealed that ultrasound (US) mediates Ca2+ influx via the synergistic action of Piezo1 and Dkk3, thereby suppressing inflammatory responses, augmenting cell vitality, and promoting neuronal activation. These findings strongly indicate that the neuroprotective effects of LITUS may be mediated via the Piezo1-Dkk3/Ca2+/PI3K-Akt3 pathway. Conclusions This study firstly elucidates the neuroprotective effects of LITUS on PCABI and clarifies the potential mechanism. We posit that this study broadens the evidence base supporting future research into the application of LITUS in clinical PCABI patients.