Mechanistic insights into the neuroprotective effects of low-intensity transcranial ultrasound stimulation in post-cardiac arrest brain injury: Modulation of the Piezo1-Dkk3/PI3K-Akt pathway
Shuang Xu, Lulu Gu, Banghe Bao, Qian Liu, Qiaofeng Jin, Yannan Ma, Siyi Zhou, Beibei Li, Li Xu, Guangqi Guo, Jinpiao Zhu, Kuan-Pin Su, Peng Sun
Brain, Behavior, and Immunity 2025 · 10.1016/j.bbi.2025.03.027
Abstract
Post-cardiac arrest brain injury (PCABI) remains a significant challenge, marked by high mortality and disability rates due to persistent neuroinflammation. This study explored the neuroprotective potential of low-intensity transcranial ultrasound stimulation (LITUS) in mitigating brain damage after cardiopulmonary resuscitation (CPR) using a murine model and in vitro assays. LITUS treatment improved 24-h survival rates and neurological recovery in cardiac arrest (CA) mice, as evidenced by behavioral assessments and reduced neurological deficit scores. Proteomic analyses revealed modulation of Piezo1-Dkk3/PI3K-Akt signaling pathway, characterized by decreased pro-inflammatory cytokines (IL-1β, IL-6, TNF-α). Mechanistic studies demonstrated that LITUS enhanced Piezo1 and Dkk3 activation, promoting calcium influx and anti-inflammatory responses. The Piezo1 antagonist GsMTx4 abrogated these effects, underscoring Piezo1's specific role. Additionally, in vitro experiments using oxygen/glucose deprivation and reoxygenation (OGD/R)-treated BV2 microglial cells confirmed that LITUS reduced inflammatory responses and enhanced cellular recovery via the Piezo1-Dkk3 axis. These findings highlight LITUS as a promising non-invasive therapeutic strategy to ameliorate PCABI by modulating neuroinflammation through the Piezo1-Dkk3/PI3K-Akt pathway. This work provides a basis for translational research and potential clinical applications in improving outcomes for CPR survivors.
Abstract via europepmc.
Exposures
Exposure 1: LITUS 500 kHz after CPR (mice) and after OGD/R (cells)
Target: brain, cultured glia — “The transducer was positioned above the wire between the eye and ear.”
Device: not reported
| Waveform | pulsed | |
|---|---|---|
| Fundamental frequency (kHz) | 500 | ✓✓✓ |
| Pulse duration (ms) | 0.4 | ✓✓✓ |
| Pulse repetition frequency (Hz) | 1,000 | ✓✓✓ |
| Duty cycle (%) | 40pulse duration × PRF gives 40% | ✓✓✓ |
| Sonication duration (s) | 0.3 | ✓✓✓⚑ |
| Free-field pressure (kPa) | not reported | |
|---|---|---|
| Free-field Isppa (W/cm²) | not reported | |
| Free-field Ispta (W/cm²) | not reported | |
| In-situ estimate | not reported | |
| In-situ pressure (kPa) | not reported | |
| In-situ Isppa (W/cm²) | not reported | |
| In-situ Ispta (W/cm²) | not reported | |
| Pressure, domain unspecified (kPa) | 300 | ✓✓✓⚑ |
0.3 MPa, 200 tone bursts, 500 kHz, PRF 1 kHz, duty cycle 40 %, sonication duration 300 ms, pulse width 400 us; stimulation interval 10 s, total stimulation 40 min starting 10 min after ROSC. In vitro, the US group received immediate US for 40 min after OGD/R.
Flags from extraction
exposures[0].timing.sonication_duration_s— 200 tone bursts at 1 kHz PRF would last 200 ms, not the stated 300 ms; text recorded as written.exposures[0].unspecified_domain.pressure_kpa— 0.3 MPa stated without saying whether free field or in situ; converted from MPa.exposures[0].target.terms— Target described only as position of transducer between eye and ear; BV2 microglia cells received US in vitro with parameters not separately restated, so cultured_glia added to one exposure.n_subjects— Group sizes given for four groups (sham 10, CPR 25, CPR+LITUS 25, CPR+LITUS+GsM 25); sham and CPR groups were not sonicated. Figure legends report n=8 per group.exposures[0].device— Driving electronics named (Rigol generator, Aigtek amplifier) but transducer maker not named.