title: The Raf-1 inhibitor GW5074 and the ERK1/2 pathway inhibitor U0126 ameliorate PC12 cells apoptosis induced by 6-hydroxydopamine.
abstract: 6-Hydroxydopamine (6-OHDA) is a widely used dopaminergic neurotoxin that leads to cell apoptosis in vivo and in vitro, and is a widely accepted experimental model of neurodegeneration in Parkinson's disease. However, the molecular mechanisms responsible for 6-OHDA-induced cell apoptosis are unclear. We found that the treatment of PC12 cells with 6-OHDA resulted in a significant decrease in cell viability and elevated apoptosis as detected by MTT assay, Hoechst 33258 staining, and flow cytometry. In addition, 6-OHDA induced a time-dependent phosphorylation of ERK1/2 at Thr-202/Tyr-204 and of Raf-1 at Ser-338, but a decreased level of Raf-1 phosphorylation at Ser-259. Phosphorylation of ERK1/2 at Thr-202/Tyr-204 and Raf-1 at Ser-338 were inhibited by the Raf-1 inhibitor GW5074, while the ERK1/2 pathway inhibitor U0126 decreased phosphorylation of ERK1/2. Furthermore, 6-OHDA-induced PC12 cells apoptosis was suppressed by GW5074 and U0126. Our results suggest that GW5074 and U0126 act as neuroprotants against 6-OHDA toxicity in PC12 cells by modulating Raf-1/ERK1/2 signaling systems.
authors.affiliation: Department of Geriatrics, the Second Affiliated Hospital of Harbin Medical University, Harbin, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Ying
authors.initials: Y
authors.lastname: Fan
authors.forename: Yi-Na
authors.initials: YN
authors.lastname: Zhang
authors.forename: Dian-Jun
authors.initials: DJ
authors.lastname: Sun
authors.forename: Song-Bin
authors.initials: SB
authors.lastname: Fu
authors.forename: Lan
authors.initials: L
authors.lastname: Ma
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Jiang
authors.forename: Can
authors.initials: C
authors.lastname: Cui
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Ding
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D006690:Bisbenzimidazole; D015153:Blotting, Western; D002070:Butadienes / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D007202:Indicators and Reagents; D007211:Indoles / Q000494:pharmacology*; D020935:MAP Kinase Signaling System / Q000187:drug effects*; D009570:Nitriles / Q000494:pharmacology*; D016627:Oxidopamine / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D016716:PC12 Cells; D010300:Parkinson Disease / Q000503:physiopathology; D010636:Phenols / Q000494:pharmacology*; D010766:Phosphorylation; D019908:Proto-Oncogene Proteins c-raf / Q000037:antagonists & inhibitors*
chemical_list: D002070:Butadienes; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D007211:Indoles; D009570:Nitriles; D010636:Phenols; C113580:U 0126; D016627:Oxidopamine; D019908:Proto-Oncogene Proteins c-raf; D006690:Bisbenzimidazole; C489251:5-iodo-3-((3,5-dibromo-4-hydroxyphenyl)methylene)-2-indolinone