title: Carvacrol protects neuroblastoma SH-SY5Y cells against Fe(2+)-induced apoptosis by suppressing activation of MAPK/JNK-NF-κB signaling pathway.
abstract: OBJECTIVE
  Carvacrol (2-methyl-5-isopropylphenol), a phenolic monoterpene in the essential oils of the genera Origanum and Thymus, has been shown to exert a variety of therapeutic effects. Here we examined whether carvacrol protected neuroblastoma SH-SY5Y cells against Fe(2+)-induced apoptosis and explored the underlying mechanisms.
  
  
  METHODS
  Neuroblastoma SH-SY5Y cells were incubated with Fe(2+) for 24 h, and the cell viability was assessed with CCK-8 assay. TUNEL assay and flow cytometric analysis were performed to evaluate cell apoptosis. The mRNA levels of pro-inflammatory cytokines and NF-κB p65 were determined using qPCR. The expression of relevant proteins was determined using Western blot analysis or immunofluorescence staining.
  
  
  RESULTS
  Treatment of SH-SY5Y cells with Fe(2+) (50-200 μmol/L) dose-dependently decreased the cell viability, which was significantly attenuated by pretreatment with carvacrol (164 and 333 μmol/L). Treatment with Fe(2+) increased the Bax level and caspase-3 activity, and decreased the Bcl-2 level, resulting in cell apoptosis. Furthermore, treatment with Fe(2+) significantly increased the gene expression of IL-1β, IL-6 and TNF-α, and induced the nuclear translocation of NF-κB. Treatment with Fe(2+) also significantly increased the phosphorylation of p38, ERK, JNK and IKK in the cells. Pretreatment with carvacrol significantly inhibited Fe(2+)-induced activation of NF-κB, expression of the pro-inflammatory cytokines, and cell apoptosis. Moreover, pretreatment with carvacrol inhibited Fe(2+)-induced phosphorylation of JNK and IKK, but not p38 and ERK in the cells.
  
  
  CONCLUSIONS
  Carvacrol protects neuroblastoma SH-SY5Y cells against Fe(2+)-induced apoptosis, which may result from suppressing the MAPK/JNK-NF-κB signaling pathways.
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Zhen-wen
authors.initials: ZW
authors.lastname: Cui
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Zheng-xing
authors.initials: ZX
authors.lastname: Xie
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Bao-feng
authors.initials: BF
authors.lastname: Wang
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Zhi-hong
authors.initials: ZH
authors.lastname: Zhong
authors.affiliation: Neuroscience and Neuroengineering Research Center, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China.
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Chen
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Yu-hao
authors.initials: YH
authors.lastname: Sun
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Qing-fang
authors.initials: QF
authors.lastname: Sun
authors.affiliation: Department of Neurology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.|Neuroscience and Neuroengineering Research Center, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China.
authors.forename: Guo-yuan
authors.initials: GY
authors.lastname: Yang
authors.affiliation: Department of Neurosurgery, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
authors.forename: Liu-guan
authors.initials: LG
authors.lastname: Bian
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002413:Cations, Divalent / Q000633:toxicity; D045744:Cell Line, Tumor; D000081007:Cymenes; D006801:Humans; D007501:Iron / Q000633:toxicity*; D048670:MAP Kinase Kinase 4 / Q000276:immunology*; D020928:Mitogen-Activated Protein Kinases / Q000276:immunology*; D039821:Monoterpenes / Q000627:therapeutic use*; D016328:NF-kappa B / Q000276:immunology*; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D002413:Cations, Divalent; D000081007:Cymenes; D039821:Monoterpenes; D016328:NF-kappa B; D018696:Neuroprotective Agents; C073316:carvacrol; D007501:Iron; D020928:Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4