Effects of low-dose lipopolysaccharide on up-regulating Nrf2 expression and reducing ROS in PC12 cells
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Effects of low-dose lipopolysaccharide on up-regulating Nrf2 expression and reducing ROS in PC12 cells
Journal of Army Medical UniversityVol. 33, Issue 5, Pages: 515-518(2011)
作者机构:
重庆医科大学附属第一医院骨科
作者简介:
基金信息:
DOI:
CLC:R338.2;R392.32;R741.02
Published:2011,
Published Online:15 March 2011,
稿件说明:
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ZHU FENGCHEN, JIANG DIANMING, LI WEICHAO, et al. Effects of low-dose lipopolysaccharide on up-regulating Nrf2 expression and reducing ROS in PC12 cells. [J]. Journal of army medical university, 2011, 33(5): 515-518.
DOI:
ZHU FENGCHEN, JIANG DIANMING, LI WEICHAO, et al. Effects of low-dose lipopolysaccharide on up-regulating Nrf2 expression and reducing ROS in PC12 cells. [J]. Journal of army medical university, 2011, 33(5): 515-518.DOI:
Effects of low-dose lipopolysaccharide on up-regulating Nrf2 expression and reducing ROS in PC12 cells
Objective To study the effects of low-dose lipopolysaccharide (LPS) on nuclear factor-erythroid 2-related factor-2 (Nrf2) expression and reactive oxygen species (ROS) level in PC12 cells. Methods LPS solutions with concentrations of 0.01
0.025
0.05
0.1
0
.25
0.5 and 1 μg/μl were used to treat PC12 cells for 24 h
and then cell viability was tested by MTT assay to determine the maximum safe dose. PC12 cells treated by LPS of the determined dose were subjected to detection of Nrf2 mRNA and protein expression in whole cell and Nrf2 protein expression in cytoplasm and nuclei through RT-PCR and Western blotting. The average fluorescence intensity of ROS in the cells was tested by flow cytometry as well. Results LPS with a concentration lower than 0.1 μg/μl had no obvious toxicity to PC12 cells (
P
>0.05). In PC12 cells treated by 0.1 μg/μl LPS for 24 h
the expression of Nrf2 mRNA and protein in whole cell increased gradually. During the process
there were significant differences for mRNA expression at 18 h as well as mRNA and total protein expression at 24 h
as compared with those of the control group (
P
<
0.01). In PC12 cells treated by 0.1 μg/μl LPS for 1
3
6 and 12 h
the Nrf2 protein expression in cytoplasm decreased gradually and then recovered slightly
with the lowest expression at 6 h
as compared with that of the control group (
P
<
0.01). The Nrf2 protein expression in the nuclei increased
with a significant difference from that of the control group (
P
<
0.01)
and showed an expression peak at 3 h. In PC12 cells treated by 0.1 μg/μl LPS for 8 h
the average fluorescence intensity of ROS decreased significantly (
P
<
0.01). Conclusion Low-dose LPS can promote the transcription
translation and nuclear translocation of Nrf2 in PC12 cells with time dependence and reduce ROS level
and probably participates in stress tolerance.
关键词
脂多糖PC12细胞NF-E2相关因子2保护机制活性氧
Keywords
lipopolysaccharidePC12 cellNF-E2-related factor 2protection mechanismreactive oxygen species
Department of Anesthesiology, Second Affiliated Hospital, Army Military Medical University (Third Military Medical University)
Department of Neurosurgery, the First Affiliated Hospital of Chongqing Medical University
College of Bioengineering, Chongqing University
Department of Neurobiology, Chongqing Municipal Key Laboratory of Neurobiology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University)
Department of Military Cognitive Psychology, Faculty of Medical Psychology, Army Medical University (Third Military Medical University)