title: [The role of neuroglobin in oxygen-glucose deprivation and reoxygenation-induced mitochondrial depolarization and reactive oxygen species production in SH-SY5Y cells].
abstract: Objective:To investigate the role of neuroglobin (NGB) in oxygen-glucose deprivation and reoxygenation (OGD/R) induced mitochondrial depolarization and reactive oxygen species (ROS)production in a human neuroblastoma cell line (SH-SY5Y).Methods:SH-SY5Y cells were transfected with lentivirus to establish a stable cell line of NGB knockdown (KD). After treated with OGD/R, cells were collected at different time points to analyze NGB mRNA and protein levels. Furthermore, cells were stained with JC-1 and DCFH-DA to evaluate mitochondrial depolarization and ROS production by inverted fluorescence microscope. Also, to determine the neurotoxicity, we measured the lactate dehydrogenase(LDH)level in the cell culture medium.Results:After the treatment of OGD/R, the NGB mRNA and protein started to elevate and peak at 4 h and 8 h (2.04±0.35 fold, 1.69±0.18 fold). Compared with the vector group, NGB KD group had much more mitochondrial depolarization [JC-1 red/green (1.10±0.10) vs (1.46±0.11),P<0.05] and ROS production [DCFH-DA fluorescence (36.30±5.32) vs (16.26±2.97),P<0.05]. Furthermore, NGB KD groups had a higher level of LDH release [(63.42±6.14)%vs (49.65±5.09)%,P<0.05].Conclusions:NGB plays an important role in the homeostasis of mitochondria. Knockdown of NGB results in increased mitochondrial depolarization, ROS production and neurotoxicity under hypoxia circumstances.
authors.affiliation: Department of Critical Care Medicine, Xiangya Hospital, Central South University, Changsha 410008, China.
authors.forename: S Y
authors.initials: SY
authors.lastname: Deng
authors.forename: Y H
authors.initials: YH
authors.lastname: Ai
authors.forename: L N
authors.initials: LN
authors.lastname: Zhang
authors.forename: L
authors.initials: L
authors.lastname: Wu
authors.forename: C X
authors.initials: CX
authors.lastname: Chen
authors.forename: Y M
authors.initials: YM
authors.lastname: Wang
authors.forename: Z Y
authors.initials: ZY
authors.lastname: Liu
authors.forename: L
authors.initials: L
authors.lastname: Huang
authors.forename: Q Y
authors.initials: QY
authors.lastname: Peng
mesh_terms: D002478:Cells, Cultured; D005452:Fluoresceins; D005914:Globins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D005947:Glucose / Q000172:deficiency* / Q000378:metabolism / Q000494:pharmacology*; D006706:Homeostasis / Q000187:drug effects*; D006801:Humans; D000860:Hypoxia / Q000473:pathology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000078787:Neuroglobin; D010100:Oxygen / Q000378:metabolism / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D014162:Transfection
chemical_list: D005452:Fluoresceins; D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; C029569:diacetyldichlorofluorescein; D005914:Globins; D005947:Glucose; D010100:Oxygen