The role of neuroglobin in oxygen-glucose deprivation and reoxygenation-induced mitochondrial ;depolarization and reactive oxygen species production in SH-SY5Y cells
10.3760/cma.j.issn.0578-1426.2017.01.011
- VernacularTitle:脑红蛋白在氧糖剥夺-复氧复糖诱导人神经母细胞瘤细胞线粒体膜电位去极化及活性氧生成中的作用
- Author:
Songyun DENG
;
Yuhang AI
;
Lina ZHANG
;
Long WU
;
Caixia CHEN
;
Yimin WANG
;
Zhiyong LIU
;
Li HUANG
;
Qianyi PENG
- Keywords:
Neuroglobin;
Membrane potential,mitochondrial;
Reactive oxygen species;
Lactate dehydrogenase;
Human neuroblastoma cells;
Oxygen glucose deprivation
- From:
Chinese Journal of Internal Medicine
2017;56(1):44-48
- CountryChina
- Language:Chinese
-
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.
