title: Protective effect of calycosin-7-O-β-D-glucopyranoside against oxidative stress of BRL-3A cells induced by thioacetamide.
abstract: BACKGROUND
  Calycosin-7-O-β-D-glucopyranoside (CG) is a natural isoflavone found in traditional Chinese medicines Astragali Radix (AR).
  
  
  OBJECTIVE
  Calycosin-7-O-β-D-glucopyranoside, an isoflavone isolated from AR, has been found to have potent antioxidantive effects. This study was designed to investigate whether CG prevents oxidative stress induced by thioacetamide (TAA).
  
  
  METHODS
  BRL-3A cells were pretreated with different concentrations of CG (10, 20, 40 mg/mL) for 12 h and then exposed to 0.18 mol/L TAA for 2 h. The cell viability were examined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium assay, total antioxidant capacity, malondialdehyde (MDA) and the activity of antioxidant enzymes, including catalase, glutathione peroxidase and superoxide dismutase were determined by microplate method. Reactive oxygen species (ROS) generation was quantified by the 2',7'-dichlorofluorescin-diacetate method. Protein and mRNA expression of CYP2E1 were determined by western blotting and real-time PCR.
  
  
  RESULTS
  The cell oxidative stress was significantly increased after 2 h of TAA exposure. Pretreatment of BRL-3A cells with CG significantly increased the activities of antioxidant enzymes, scavenged ROS and reduced MDA production. CG decreased the expression of CYP2E1, and ultimately decreased TAA-induced BRL-3A cells oxidative stress.
  
  
  CONCLUSIONS
  Calycosin-7-O-β-D-glucopyranoside has a protective effect against TAA-induced oxidative stress in BRL-3A cells, and that the underlying mechanism involves in scavenging of ROS and the modulating expression of CYP2E1.
authors.affiliation: University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, China.
authors.forename: Li
authors.initials: L
authors.lastname: Jian
authors.affiliation: University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Xin
authors.affiliation: University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, China.
authors.forename: Ma
authors.initials: M
authors.lastname: Yufang
authors.affiliation: University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal Healthcare in Fujian Province, Fujian Agriculture and Forestry University, Fuzhou, China.
authors.forename: Huang
authors.initials: H
authors.lastname: Yifan
keywords: BRL-3A cell; CYP2E1; calycosin-7-O-β-D-glucopyranoside; oxidative stress; thioacetamide