To investigate the mechanism of Dazao (Ziziphi Jujubae Fructus) in reducing Fuzi (Aconiti Lateralis Radix)-induced neurotoxicity based on network pharmacology and animal experiments.

The toxic components of Fuzi (Aconiti Lateralis Radix) and active constituents of Dazao (Ziziphi Jujubae Fructus) were screened through literature retrieval and database searches. Potential targets of toxic components were predicted using Swiss Target Prediction and PharmMapper databases, followed by construction of a comprehensive target library. A disease target library was established by retrieving “nerve toxicity” and “toxic encephalopathy”-related genes from biomedical databases. The intersection targets between component-related and disease-related targets were subsequently subjected to protein-protein interaction (PPI) network analysis and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment. Comparative analysis of the identified detoxification pathways [from Dazao (Ziziphi Jujubae Fructus)] and neurotoxicity pathways [from Fuzi (Aconiti Lateralis Radix)] was performed to predict the underlying detoxification mechanisms. Using SD rats as the animal model, a quantitative real-time polymerase chain reaction (qRT-PCR) was employed to investigate the effects of Fuzi (Aconiti Lateralis Radix) and Dazao (Ziziphi Jujubae Fructus) combination therapy on mRNA expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and hypoxia inducible factor-1α(HIF-1α) in the brain tissues of the rats.

Through network pharmacology analysis, it identified 21 potential toxic components and 1169 potential toxic targets in Fuzi (Aconiti Lateralis Radix), including 570 neurotoxic targets. There were 29 active components in Dazao (Ziziphi Jujubae Fructus) with 680 corresponding targets. Venn analysis identified 76 overlapping targets as the potential detoxification targets through which Dazao (Ziziphi Jujubae Fructus) counteracted Fuzi (Aconiti Lateralis Radix)-induced neurotoxicity. PPI network pharmacology identified quercetin, saponins and specific alkaloids as the primary mediators of Dazao (Ziziphi Jujubae Fructus) detoxification effects against Fuzi (Aconiti Lateralis Radix) neurotoxicity. The KEGG pathway analysis identified enrichment in 171 pathways, primarily involving signaling pathways such as the PI3K-AKT signaling pathway and HIF-1 signaling pathway. In vivo studies revealed that Fuzi (Aconiti Lateralis Radix) disrupted redox homeostasis in the rat brain, promoting oxidative stress, whereas co-administration with Dazao (Ziziphi Jujubae Fructus) effectively counteracted these pro-oxidative effects, q RT-PCR experiment results demonstrated that, compared with those of the control group, the mRNA levels of PI3K, AKT and HIF-1α in the cerebral cortex and hippocampal tissues of the rats in the Fuzi (Aconiti Lateralis Radix) group were significantly increased. Compared with those in the Fuzi (Aconiti Lateralis Radix) group, the mRNA expression levels of PI3K, AKT and HIF-1α in the cerebral cortex and hippocampus of rats in the Fuzi (Aconiti Lateralis Radix)-Dazao (Ziziphi Jujubae Fructus) combination group were significantly decreased.

The neuroprotective effect of Dazao (Ziziphi Jujubae Fructus) against Fuzi (Aconiti Lateralis Radix)-induced neurotoxicity is mediated through modulation of the PI3K/AKT/HIF-1 signaling pathway, thereby alleviating Fuzi (Aconiti Lateralis Radix)-triggered oxidative stress damage in rat brain tissues. These findings provide a novel perspective for network pharmacology research on toxic traditional Chinese herbs and offer new insights into the mechanism of toxicity reduction through herbal compatibility.