Abstract: Objective To explore the toxic target organs and potential mechanism of periplocin based on a zebrafish model. Methods Zebrafish(4 days post fertilization,4 dpf) were exposed to different concentrations of periplocin solution for 24 h (the low concentration was 1 μg·mL^-1and the high concentration was 1.5 μg·mL^-1), and the fatality rate of periplocin for zebrafish was calculated. After twenty-four hours of exposure to a sub-lethal dose, the hepatotoxicity of periplocin was evaluated based on changes of areas of the liver, aeridine orange staining, liver histopathologic sections, and changes in alanine aminotransferase (ALT) and aspartic aminotransferase (AST) activities. Furthermore, the potential mechanism of periplocin hepatotoxicity was predicted via network pharmacology and molecular docking techniques before being verified by real-time PCR. Results The sublethal dose of periplocin for zebrafish was 1.612 6 μg·mL^-1. Under sub-lethal dose exposure, periplocin treatment could result in reduced liver areas of zebrafish and obvious apoptosis in the liver compared with the control group. Histopathological section results showed that periplocin exposure induced loose and disordered arrangement of liver cells as well as obvious vacuoles. In addition, periplocin could significantly increase the activities of ALT and AST in zebrafish, which proved the significant hepatotoxicity of periplocin in zebrafish. Network pharmacology indicated that periplocin-induced hepatictoxicity was related to the 21 potential targets and the top five targets of PPI protein interaction network were STAT3, HSP90AA1, HIF1A, NOS3 and MTOR. KEGG results showed that periplocin hepatotoxicity might be related to HIF-1 signaling pathway, PI3K/Akt signaling pathway, and calcium signaling pathway, as was confirmed by RT-PCR. Conclusion Periplocin-induced hepatotoxicity is associated with the disturbance of HIF-1 and PI3K/Akt signaling pathways.

Key words: periplocin, hepatotoxicity, zebrafish, mechanism