Yiqi Huoxue formula attenuates cerebral ischemia-reperfusion injury by targeting miR-29b-3p: Integrated bioinformatics and experimental validation.

This study aimed to systematically clarify the therapeutic effects and underlying mechanisms of Yiqi Huoxue Formula (YQHXF) against cerebral ischemia-reperfusion injury (CIRI) by integrating bioinformatics, network pharmacology, and in vitro/in vivo experiments. Key active compounds and targets of YQHXF were predicted by network pharmacology. Autophagy-related differentially expressed microRNAs (miRNAs) in CIRI were screened from GEO database; target genes of candidate miRNA were verified by dual-luciferase reporter assay. Rat middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced SH-SY5Y cell models were established to simulate CIRI. Neurological function, infarct volume, and pathological morphology were evaluated. Autophagosomes were observed by transmission electron microscopy. LC3B and p62 expression was detected by immunofluorescence. Cell viability and apoptosis were measured by CCK-8 assay and flow cytometry, respectively. Expression of target miRNA and PTEN-PI3K/AKT/mTOR pathway was determined by Western blotting and RT-qPCR. YQHXF significantly improved neurological function, reduced infarct volume, alleviated brain pathological damage, and inhibited autophagic activity in MCAO/R rats. In SH-SY5Y cells, YQHXF enhanced cell viability, improved morphology, reduced apoptosis, and suppressed autophagy. Bioinformatics identified miR-29b-3p as a key autophagy-regulating miRNA in CIRI, with PTEN as its direct target. Network pharmacology showed PIK3CA, MAPK8, and BCL2L1 as core targets of YQHXF. YQHXF upregulated miR-29b-3p, downregulated PTEN, and activated PI3K/AKT/mTOR pathway; miR-29b-3p antagonist reversed YQHXF's therapeutic effects in vivo. YQHXF exerts neuroprotective effects against CIRI by targeting miR-29b-3p, providing a basis for its further research and clinical application.