Banxia Baizhu Tianma decoction inhibits neuronal cell death, inflammatory injury, and blood-brain barrier disruption after SAH via the PI3K-AKT pathway.

BACKGROUND: Neuronal programmed cell death, inflammation, and blood-brain barrier (BBB) disruption drive secondary injury after subarachnoid hemorrhage (SAH). However, no multi-target therapies are currently available. Banxia Baizhu Tianma Decoction (BBTD) shows therapeutic potential for cerebral hemorrhage. This study aimed to investigate BBTD's protective mechanisms against these pathologies post-SAH. METHODS: A rat SAH model was established via endovascular perforation. Neuroprotection was assessed using behavioral tests (Foot-Fault, Rotarod, Forelimb Placing, Garcia scores, and water maze), molecular analyses (Western blot, TUNEL, immunofluorescence), BBB assays (edema, Evans Blue/FITC-dextran permeability), and inflammation evaluation (microglial polarization, cytokine ELISA). Signaling pathways were screened via mass spectrometry/network pharmacology and validated with the PI3K inhibitor LY294002. RESULTS: BBTD attenuated neuronal apoptosis by suppressing caspase-3/7/8/9, Bax, and Cytc activation; inhibited necroptosis via suppression of the RIP1/RIP3/MLKL complex; and reduced ferroptosis by downregulating ACSL4 while upregulating GPX4. BBB integrity was restored through increased ZO-1/occludin/claudin-5 expression, inhibition of MMP-3/9/13, and enhanced VASP/eNOS phosphorylation. Inflammation was mitigated via microglial M1-to-M2 polarization shift, reduced cytokine release, and NLRP3/caspase-1 downregulation. The PI3K-AKT pathway was identified as the core mechanism and functionally validated, and the top five high-affinity molecules were identified. CONCLUSION: BBTD confers neuroprotection against SAH by multi-target inhibition of programmed cell death, BBB disruption, and neuroinflammation, primarily via PI3K-AKT activation. This provides a foundation for preclinical evidence. DATA AVAILABILITY: The data from this study will be made available to qualified investigators upon reasonable inquiry.