Bufothionine induced the mitochondria-mediated apoptosis in H22 liver tumor and acute liver injury
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) (Inj A) and bufothionine (9.77 mL/kg) (Bufo A) 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, H22-tumor-bearing mice were randomly divided into five groups (n=10): normal saline group, model group, cinobufacini injection (Inj B) (5.14 mL/kg), bufothionine (8.02 mL/kg)(Bufo B) and 5-fluorouracil (5-Fu) (3.42 mL/kg).Tumors were picked out and determined with vemier 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, 90.62μg/mL).Results: Butothionine relieved CCl4-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 H22-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 and0.0464), 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.
Rui-Fang Xie Zhi-Cheng Li Pei-Pei Chen Xin Zhou
国内会议
郑州
英文
353-371
2015-10-01（万方平台首次上网日期，不代表论文的发表时间）