Puerarin Protects from Methotrexate Induced Hepatotoxicity in AML-12 Cells

Aim: The purpose of this study was to look into the effects of puerarin (PR) on methotrexate (MTX)-induced hepatotoxicity in vitro. Materials and Methods: We designed our research with four groups in the AML-12 cell line: control, PR, MTX, and PR+MTX groups. Administered concentration levels to the cell lines were determined with the MTT test. To investigate oxidative stress, the expression levels of glutathione, superoxide dismutase, and catalase were determined with quantitative real-time-polymerase chain reaction (qRT-PCR) analysis. To evaluate the role of apoptosis pathways in MTX induced hepatotoxicity and the hepatoprotective effects of PR, gene expressions of caspase 3 (Cas-3), Cas-9, apoptotic protease activating factor-1, Bcl-2, Bax, p53, second mitochondria-derived activator of caspase/direct inhibitor of apoptosis-binding protein (smac/DIABLO), topoisomerase (Top) I, and Top II were investigated with qRT-PCR. Results: MTX impaired the antioxidant defense through SOD but elevated the expression of catalase and glutathione due to an increase in free radicals. In the PR+MTX group, SOD expression increased and catalase and glutathione expression decreased compared to the MTX group. Cas-9, Apaf-1, and Top I gene expression levels were reduced in group PR. In the group of PR+MTX, PR application increased the expression of Bax, p53, and smac/DIABLO while decreasing the expression of Bcl-2, which resulted in the elimination of damaged structures by apoptosis. Conclusion: PR alleviated the hepatotoxicity caused by MTX with its antioxidant effects and positive effects on apoptosis pathways. However, different dose studies are needed because PR could not prevent double-strand damage in DNA due to MTX and there is an increase in Top I expression in the PR group.