Hepatoprotective effect of rhein against methotrexate-induced liver toxicity

The purpose of this study was to investigate the protective effect of rhein, a major metabolite of diacerein, on methotrexate (MTX)-induced hepatotoxicity and clarify the pharmacological mechanism. Rhein significantly reduced the elevation of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) caused by MTX in rat serum and improved liver morphological damage induced by MTX. Moreover, rhein increased the cell survival rate and reduced the number of apoptosis cells in MTX-treated normal human hepatocyte (L02 cells). Rhein treatment in rats up-regulated nuclear factor erythroid 2-related factor 2 (Nrf2), B-cell lymphoma-2 (Bcl-2), heme oxygenase 1 (HO-1) and glutamate-cysteine ligase catalytic subunit (GCLC), and down-regulated Bcl-2 associated x (Bax) in mRNA and protein levels. Furthermore, rhein treatment further decreased protein expression of nuclear factor-kappa B (NF-κB), tumor necrosis factor alpha (TNF-α) and cysteine aspartic acid specific protease 3 (Caspase-3), increased protein expression of B-cell lymphoma-extra large (Bcl-xl), and reduced mRNA expression of Bcl-2 homologous antagonist/killer (Bak) in MTX-treated rat liver in vivo. However, the protein expression changes of Nrf2, HO-1, GCLC, Bcl-2, Bcl-xl and Bax could be abrogated by Nrf2 antagonist brusatol. In addition, protective effect of rhein against MTX-mediated liver damage could also be suppressed by Nrf2 siRNA in L02 cells. Taken together, these findings suggested that rhein ameliorated liver damage mediated by MTX through acting on Nrf2-HO-1 pathway. NF-κB, TNF-α, Caspase-3 and Bcl-2 family were also participated in the protection. As effectively hepatoprotective ability of rhein, it would raise an important issue for patients orally receiving MTX treatment together with diacerein/rhein.