title: Berberine Induces Cell Apoptosis through Cytochrome C/Apoptotic Protease-Activating Factor 1/Caspase-3 and Apoptosis Inducing Factor Pathway in Mouse Insulinoma Cells.
abstract: OBJECTIVE
  To investigate apoptotic effects of berberine, a significant alkaloids component existing in Rhizoma coptidis, and its possible acting mechanism in insulinoma cells.
  
  
  METHODS
  Different concentrations of berberine were used to treat mouse insulinoma (MIN6) cells for various period of time. The viability and apoptosis of the cells were analyzed using methylthiazolyldiphenvl-tetrazolium bromide assay, flow cytometry and enzyme-linked immuno sorbent assay. Changes in the relating pro- and anti-apoptosis proteins were detected by western-blotting.
  
  
  RESULTS
  The half-maximal inhibitory concentration (IC50) of berberine was 5.7 μmol/L on MIN6 cells viability for 16 h. Berberine caused a 20% reduction (P<0.05) in cell number after only 4-h incubation; which reached 50% after 24 h (P<0.01). Berberine treatment for 16 h significantly increased the level of DNA fragmentation. The flow cytometry showed the apoptotic rate increased 2.9- and 4.6-fold after treating with berberine (5 μmol/L) for 8 and 16 h, while 3- and 8.7-fold after 10 μmol/L treatment for 8 and 16 h (P<0.01). Berberine treatment dramatically elevated the expression ratio of Bax to Bcl-2. Meanwhile, berberine notably increased the apoptosis-inducing factors and cytochrome C transforming from the mitochondria to the cytoplasm. Apoptotic protease-activating factor 1 (Apaf-1) was subsequently activated after cytochrome C release. Furthermore, caspase-3 and poly adenosine diphosphate-ribose polymerase were also activated to trigger apoptosis cascade.
  
  
  CONCLUSIONS
  High concentration (5 and 10 μmol/L) of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor (AIF) pathway.
authors.affiliation: School of Basic Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China.|Fraser Laboratories for Diabetes Research, Department of Medicine, RI-McGill University Health Centre, Montreal, Quebec, H4A3J1, Canada.
authors.forename: Xin
authors.initials: X
authors.lastname: Fang
authors.affiliation: Fraser Laboratories for Diabetes Research, Department of Medicine, RI-McGill University Health Centre, Montreal, Quebec, H4A3J1, Canada.|School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Xiao-Liang
authors.initials: XL
authors.lastname: Miao
authors.affiliation: Fraser Laboratories for Diabetes Research, Department of Medicine, RI-McGill University Health Centre, Montreal, Quebec, H4A3J1, Canada.
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Liu
authors.affiliation: Diabetes Research Center, Beijing University of Chinese Medicine, Beijing, 100029, China.
authors.forename: Dong-Wei
authors.initials: DW
authors.lastname: Zhang
authors.affiliation: School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.affiliation: Diabetes Research Center, Beijing University of Chinese Medicine, Beijing, 100029, China.
authors.forename: Dan-Dan
authors.initials: DD
authors.lastname: Zhao
authors.affiliation: School of Basic Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China.
authors.forename: Qian-Qian
authors.initials: QQ
authors.lastname: Mu
authors.affiliation: School of Basic Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China.
authors.forename: Na
authors.initials: N
authors.lastname: Yu
authors.affiliation: Diabetes Research Center, Beijing University of Chinese Medicine, Beijing, 100029, China.
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Mo
authors.affiliation: School of Life Science and Technology, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Hong-Ping
authors.initials: HP
authors.lastname: Yin
authors.affiliation: School of Basic Medicine, Beijing University of Chinese Medicine, Beijing, 100029, China. gaosihua1216@163.com.|Diabetes Research Center, Beijing University of Chinese Medicine, Beijing, 100029, China. gaosihua1216@163.com.
authors.forename: Si-Hua
authors.initials: SH
authors.lastname: Gao
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D053477:Apoptotic Protease-Activating Factor 1 / Q000378:metabolism; D001599:Berberine / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D007340:Insulinoma / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured
chemical_list: D051033:Apoptosis Inducing Factor; D053477:Apoptotic Protease-Activating Factor 1; D001599:Berberine; D045304:Cytochromes c; D053148:Caspase 3
keywords: Chinese medicine; apoptosis; apoptosis inducing factor pathway; berberine; cytochrome C/Apaf-1/caspase-3; mouse insulinoma cells