Angong Niuhuang Pill mitigates hypoxia/ischemia-induced impairments in neuromotor function and microglial activation in neonatal rats

Abstract

Cerebral palsy secondary to hypoxic–ischemic encephalopathy remains a leading cause of neonatal neurological disability with limited treatment options. Angong Niuhuang Pill, a traditional Chinese medicine formulation, has demonstrated neuroprotective potential. However, its efficacy and mechanisms in cerebral palsy models remain underexplored. In this study, we evaluated the therapeutic effects of Angong Niuhuang Pill in a neonatal rat model of hypoxia/ischemia-induced cerebral palsy. Magnetic resonance imaging revealed significantly reduced infarct volume in Angong Niuhuang Pill-treated animals compared with the vehicle group. Behavioral assessments showed improved negative geotaxis performance on day 14, as well as enhanced outcomes in foot fault and von Frey filament tests after day 42, indicating amelioration of motor deficits and sensory disturbances. Histological analysis revealed decreased microglial activation and reduced cell counts in the ipsilateral hemisphere. Transcriptomic sequencing and Western blot analyses confirmed the downregulation of pyroptosis-related molecules, including NOD-like receptor family pyrin domain-containing 3, gasdermin D, and the N-terminal fragment of gasdermin D (N-GSDMD), in the Angong Niuhuang Pill group. Angong Niuhuang Pill confers neuroprotection in a neonatal hypoxic-ischemic encephalopathy model of cerebral palsy by limiting infarct progression, attenuating neuroinflammation and pyroptosis, and improving long-term sensorimotor outcomes. Attenuation of the NLRP3 inflammasome pathway represents an important contributing mechanism underlying these beneficial effects.