Integrated transcriptomic and proteomic analysis of hepatotoxic effects of Venenum Bufonis in zebrafish

Venenum Bufonis (VB), a traditional Chinese medicine (TCM), is renowned for its therapeutic detoxification, pain relief, and cognitive enhancement effects. VB has been classified as a toxic TCM in medical literature, and its clinical usage is currently subject to several limitations. However, the toxicological characteristics of VB and underlying mechanisms remain unclear.

We conducted a comprehensive assessment to confirm the target organs affected by VB using the zebrafish model. Subsequently, network pharmacology, transcriptomic and proteomic analyses were performed to explore the associated mechanisms, with the aim of providing a basis for its clinical application.

VB exhibited dose-dependent toxic effects on zebrafish, particularly causing gross morphological abnormalities in the liver along with aggravated hepatocyte apoptosis. Pericardial edema and an enlarged atrioventricular septum were also observed. The combined analyses revealed significant alterations induced by VB in gene expression enriched in multiple pathways and biological processes. Importantly, TLR4/RIPK2/NF-κB and Wnt signaling-mediated inflammation, fibrosis, and apoptosis were identified as the key functional signaling pathways underlying VB-mediated liver toxicity.

Our results present robust and direct evidence of the hepatotoxic effects induced by VB in zebrafish, while also providing novel insights into the molecular pathways involved. These results establish a solid theoretical foundation for the appropriate clinical application of VB.