title: Medicated rat serum containing Gengnianchun decoction reduces apoptosis of pheochromocytoma cells insulted by amyloid beta protein.
abstract: OBJECTIVE
  To investigate the effects of medicated rat serum containing Gengnianchun (GNC) decoction and its protection to pheochromocytoma cells (PC12 cells) from amyloid beta (Abeta)(25-35)-insulted apoptosis and to find the possible mechanism.
  
  
  METHODS
  Medicated rat serum was prepared by administering ovariectomized Sprague-Dawley (SD) rats with GNC decoction. The effects of medicated rat serum on viability of PC12 cells were evaluated by cell counting kit-8 (CCK-8) assay. The PC12 cells were cultured with different doses of Abeta(25-35) to induce a model of Alzheimer's disease in vitro. Then, the protective effects of medicated rat serum on Abeta(25-35)-insulted PC12 cells were evaluated by using CCK-8 assay to detect the cell viability, using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) flow cytometry to detect cell apoptosis rate and using Western blotting assay to analyze the expressions of Bcl-2, Bax and active caspase-3 proteins.
  
  
  RESULTS
  PC12 cells cultured with 20% medicated rat serum containing GNC decoction for 24 h or 48 h had higher viability than those cultured with normal culture medium (P<0.05). After 24- or 48-hour treatment of different concentrations of Abeta(25-35), cell viabilities were all decreased as compared with normal medium (P<0.05). Cells underwent apoptosis, which showed the neurotoxicity of Abeta(25-35). The cell apoptosis induced by Abeta 25-35 was significantly decreased in PC12 cells which were pretreated with 20% medicated rat serum or nerve growth factor (NGF) according to CCK-8 assay and Annexin V-FITC/PI flow cytometry (P<0.05). The ratio of Bax expression to Bcl-2 expression and the expression of active caspase-3 were decreased in the cells treated with medicated serum or NGF as compared with the cells cultured with Abeta(25-35) only.
  
  
  CONCLUSIONS
  The GNC-medicated rat serum at concentration of 20% can promote viability of Abeta(25-35)-insulted PC12 cells and decrease the cell apoptosis by regulating the expressions of Bcl-2, Bax and active caspase 3.
authors.affiliation: Department of Integrated Traditional Chinese and Western Medicine, Fudan University, Shanghai 200011, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Wen-jiang
authors.initials: WJ
authors.lastname: Zhou
authors.forename: Fan-gui
authors.initials: FG
authors.lastname: Zhao
authors.forename: Da-jin
authors.initials: DJ
authors.lastname: Li
authors.forename: Wen-jun
authors.initials: WJ
authors.lastname: Wang
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism*; D016229:Amyloid beta-Peptides / Q000633:toxicity*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000494:pharmacology*; D016716:PC12 Cells / Q000187:drug effects; D010446:Peptide Fragments / Q000633:toxicity*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D044967:Serum; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D016229:Amyloid beta-Peptides; C494828:Bax protein, rat; D004365:Drugs, Chinese Herbal; D010446:Peptide Fragments; D019253:Proto-Oncogene Proteins c-bcl-2; C074636:amyloid beta-protein (25-35); D051028:bcl-2-Associated X Protein; C505410:Casp3 protein, rat; D053148:Caspase 3