title: Protective effect of 7-difluoromethoxy-5,4'-Di-hydroxyl isoflavone against             the damage induced by glutamate in PC12 cells.
abstract: 7-difluoromethoxy-5,4'-Di-hydroxyl isoflavone (dFGEN), prepared by the difluoromethylation             of genistein, is an active chemical entity. In this study, our main purpose was             to investigate whether dFGEN had an effect on glutamate-induced apoptosis in cultured             PC12 cells. The PC12 cells were treated with different glutamate concentrations             for 24 h in vitro. The PC12 cells impaired by glutamate were used as the cell             model of excitability. Cells were incubated for 30 min with genistein, dFGEN,             vitamin E, and exposed to 10 mM glutamate for 24 h. Cell morphology was observed             by light microscopy. The growth and proliferation of PC12 cells were detected             by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Cell             apoptosis was determined by flow cytome-try (FCM) with propidium iodide (PI) staining.             The activity of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and the             content of malondialdelyde (MDA) were measured by kits, respectively. Acridine             orange (AO) staining was used to detect characteristics of cell apoptosis. When             PC12 cells were incubated with glutamate for 24 h, cells appeared to have significant             changes in shape. The cellular viability was reduced and the apoptotic rate was             increased. The levels of LDH and the content of MDA were increased. The activity             of SOD was decreased. After PC12 cells were pretreated with dFGEN, dFGEN significantly             improved cell morphology, cell growth and proliferation, suppressed apoptosis             of cells, reduced the release of LDH, improving SOD activity and decreased MDA             content in a concentration-dependent manner. AO staining displayed that apoptosis             was decreased. These results suggested that dFGEN has a protective effect against             glutamate-induced damage in PC12 cells. dFGEN seemed to have a better protective             effect than the lead compound genistein in a concentration-dependent manner. The             mechanism of protective effect of dFGEN may be mainly related to its antioxidative             activity.
authors.affiliation: College of Medicine, Hunan Normal University, Changsha, Hunan 410006, PR China. fengxing01@hotmail.com
authors.forename: Xing
authors.initials: X
authors.lastname: Feng
authors.forename: Qin
authors.initials: Q
authors.lastname: Zhou
authors.forename: Chong
authors.initials: C
authors.lastname: Liu
authors.forename: Mei-Ling
authors.initials: ML
authors.lastname: Tao
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: Cao
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Zhong
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D005434:Flow Cytometry; D018698:Glutamic Acid / Q000494:pharmacology*; D007529:Isoflavones / Q000494:pharmacology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D019636:Neurodegenerative Diseases / Q000188:drug therapy / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016716:PC12 Cells; D051381:Rats; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: C578675:7-difluoromethoxy-5,4'-dihydroxylisoflavone; D007529:Isoflavones; D018696:Neuroprotective Agents; D018698:Glutamic Acid; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase