The Screening of Major Hepatotoxic Components of Tripterygium wilfordii Hook. F. Based on Hepatotoxic Injury Patterns

Background: Tripterygium wilfordii Hook. F. (TwHF), a traditional Chinese medicine, is widely used in treatment of rheumatoid arthritis (RA), etc.. Due to multiorgan toxicity, particularly hepatotoxicity, the application of TwHF is restricted. In order to clarify the hepatotoxic substances, zebrafishes, hepatocytes and macrophages were used for screening based on hepatotoxic injury patterns. It provided a basis for further hepatotoxic mechanism of TwHF. Methods: : First, 12 compounds were selected according to the chemical categories of TwHF. The fluorescence area and fluorescence intensity of zebrafishes liver were observed and calculated. The viability of two hepatocyte lines were detected by CCK 8 . TNF-α and IL-1β mRNA expression of BMDM was used to evaluate macrophage activation, a factor of potential indirect hepatotoxicity. Finally, hepatotoxic characteristics of 4 representative components were verified by mice in vivo . Results: : Parthenolide, Triptolide, Triptonide, Triptobenzene H, Celastrol, Demethylzeylasteral, Wilforlide A, Triptotriterpenic acid A and Regelidine can significantly reduce the fluorescence area and fluorescence intensity of zebrafish liver. The viability of L-02 or AML-12 was significantly inhibited by Parthenolide, Triptolide, Triptonide, Celatrol, Demethylzeylasteral, Triptotriterpenic acid A. Parthenolide, Triptolide, Triptonide, Celatrol, Demethylzeylasteral and Triptobenzene H significantly increased TNF-α and IL-1β mRNA levels of BMDM, while Triptophenolide, Hypodiolide and Wilforine significantly reduced. Triptotriterpenic acid A, Celastrol and Triptobenzene H at dose of 10 mg/kg significantly increased the levels of mice serum ALT and AST, and aggravated liver inflammation. Conclusions: : Parthenolide, Triptolide, Triptonide, Celatrol, Demethylzeylasteral, Triptotriterpenic acid A and Triptobenzene H might be main hepatotoxic components of TwFH. Among them, Triptotriterpenic acid A only presented a direct hepatotoxicity. Triptobenzene H showed indirect liver damage by activating macrophages. Parthenolide, Triptolide, Triptonide, Celatrol, Demethylzeylasteral could directly and indirectly cause liver injury.