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Background  Bufothionine is an alkaloid in Cinobufacini ( Huachansu ). This study aims to investigate the effects of bufothionine on liver tumors and acute liver injury.    Methods  In the hepatoprotective experiment, fifty rats were randomly divided into five groups (n = 10): normal saline group, model group, compound glycyrrhizin injection (9.14 mL/kg); cinobufacini injection (3.42 mL/kg) (InjA) and bufothionine (9.77 mL/kg) (BufoA) group. Liver weight indices were recorded to judge the degree of liver swelling, hematoxylin and eosin (H&E) staining of liver tissues was carried out to observe liver histological morphology injury and biochemical indicators including aspartate aminotransferase (AST); alanine aminotransferase (ALT); alkaline phosphatase (ALP); and total bilirubin (TBIL) were determined by modular auto-analyzer. In anti-tumor experiment, H 22 -tumor-bearing mice were randomly divided into five groups (n = 10): normal saline group, model group, cinobufacini injection (InjB) (5.14 mL/kg), bufothionine (8.02 mL/kg) (BufoB) and 5-fluorouracil (5-Fu) (3.42 mL/kg). Tumors were picked out and determined with vernier calipers. Histological morphology of tumors was observed by H&E staining. In SMMC-7721 cells, expressions of proteins related to mitochondria-mediated apoptosis pathway including Bcl-2, Bax, caspase-3, caspase-9, cyto-c, Bid, and p53 were analyzed by western blotting at low, medium, high concentrations of bufothione (3.62 μg/mL, 18.12 μg/mL,90.62 μg/mL).    Results  Butothionine relieved CCl 4 -induced liver morphology, decreased the level of ALT ( P  =2.46 × 10 -2 ) and expressed tendency to decrease other biochemical markers including AST, ALP and TBIL. Butothionine could also promote necrosis of tumor tissue in H 22 -tumor-bearing mice and restrained tumor growth with 65.16% inhibition rate. Its mechanism might relate to up-regulation of p53 (at low, mediate and high concentration, corresponding  P  values were 0.142, 0.0257, 0.0162), caspase-3 ( P  = 0.246, 0.0267 and 0.0236), cyto-c ( P  = 0.276, 0.0343 and 0.0429), Bid ( P  = 0.0125, 0.0395 and 0.0132) and Bax ( P  = 0.563, 0.0492 and 0.0357) in a dose-dependent manner, down-regulation of Bcl-2 expression ( P  = 0.0232, 0.0178 and 0.0464), but had no significant effects on caspase-9 ( P  = 0.253, 0.147 and 0.287).    Conclusion  Bufothionine induced the proteins for the mitochondria-mediated apoptosis that inhibits liver tumors and protects the liver against acute injury.

Bufothionine induced the mitochondria-mediated apoptosis in H22 liver tumor and acute liver injury