Shexiang Huayu Xingnao Granule Alleviates Intracerebral Hemorrhage via PI3K/AKT‐Mediated Neuroinflammation Regulation: Integrated Network Pharmacology and Experimental Validation

OBJECTIVE: Employing network pharmacology prediction combined with experimental validation, this research elucidated the underlying mechanisms of Shexiang Huayu Xingnao granules (SXHY) against intracerebral hemorrhage (ICH). METHODS: Potential targets and related signaling pathways of SXHY acting on ICH were predicted through network pharmacology. An autologous blood injection-induced rat ICH model was established, with simvastatin as the positive control drug. The therapeutic effects of SXHY were evaluated using neurological function scores, brain water content, superoxide dismutase (SOD), catalase, malondialdehyde activity, and brain tissue hemorrhage area. Mechanism studies used a lipopolysaccharide-stimulated BV-2 microglial cell model, assessing inflammatory mediator expression profiles and PI3K/AKT pathway modulation through RT-qPCR and Western blot analysis. RESULTS: Network pharmacology analysis revealed 360 common SXHY-ICH targets, with 10 core targets (including TNF, AKT1, IL-6, and IL-1β) significantly enriched in the PI3K/AKT, AGE-RAGE, and MAPK pathways; molecular docking further demonstrated potent binding of the key component stigmasterol to TNF-α (binding energy: -9.125 kcal/mol) and IL-6 (binding energy: -8.57 kcal/mol). Animal experiments showed within-group improvements in neurological function from day 2 to day 3 post-ICH in SXHY groups. The observed proportions of animals with improved neurological scores were 88.9% (low-dose), 83.3% (high-dose), 70.0% (model), and 62.5% (simvastatin). However, no statistically significant between-group differences were observed at day 3 (p > 0.05). It also reduced brain edema (high dose: 78.0 ± 0.7% vs. MODEL: , p < 0.05); meanwhile, it significantly increased SOD activity (high dose: 68.7 ± 11.3 U/mg protein vs. model: 37.3±14.1 U/mg protein, p < 0.01). Cell experiments further confirmed that SXHY mediates anti-inflammatory activity through modulation of PI3K/AKT signaling, including reduced p-PI3K/GAPDH (p < 0.01), downregulation of proinflammatory cytokines TNF-α/IL-6/IL-1β mRNA (p < 0.01), and upregulation of anti-inflammatory cytokine IL-10 mRNA (p < 0.05). CONCLUSION: SXHY may alleviate post-ICH brain injury by inhibiting neuroinflammation throu modulation of the PI3K/AKT pathway. The results provide new mechanistic evidence for traditional Chinese medicine interventions against ICH.