Role of NRF2 signaling pathway in trichloromethane-induced oxidative stress in L02 cells
- Author:
TAN Shi ying ZHONG Yuan fang QIU Yi bing ZOU Zhi hui YU ri an
- Publication Type:Journal Article
- Keywords:
Trichloromethane Nuclear factor erythroid 2 related factor 2 Signaling pathway Oxidative stress Oxidative ； damage L02 cells
- From:
China Occupational Medicine
2022;49(05):530-
- CountryChina
- Language:Chinese
-
Abstract:
Objective - （ ）
To investigate the effects of nuclear factor erythroid 2 related factor 2 NRF2 on the oxidative stress
（ ） Methods ） ，，
induced by trichloromethane TCM in human normal hepatocyte L02 cells. i L02 cells were stimulated with 1 2
， ， ， （ ），
4 8 12 16 and 20 mmol/L TCM solution dissolved in dimethyl sulfoxide and the control group and blank group were set
， - ，
up. After culturing for 24 hours the cell viability was detected by CCK 8 colorimetric method and the concentration of TCM
） -， -
stimulation was screened. ii L02 cells in logarithmic growth phase were randomly divided into control group and low medium
- ， ， ，
and high dose groups. After 24 hours of exposure to 0 4 8 and 12 mmol/L TCM the cells were collected. The activity of
（ ）， （ ）， （ - ） （ ）
superoxide dismutase SOD catalase CAT glutathione peroxidase GSH Px and the level of malondialdehyde MDA
NRF2， - （HO-1），
were detected by colorimetric analysis. The mRNA expression levels of heme oxygenase 1 glutamate cysteine
（GCLC） （） （NQO1） -
ligase catalytic subunit and NAD P H quinone dehydrogenase 1 were detected by real time fluorescence
， - ，
polymerase chain reaction. The protein levels of NRF2 HO 1 GCLC and NQO1 were detected by Western blotting.Results ） ， ， ， ，
i When the concentration of TCM was 4 8 12 16 and 20 mmol/L the survival rate of L02 cells decreased
（ P ） ， ，
significantly compared with the control group all <0.05 . The concentration of 0 4 8 and 12 mmol/L were selected as the
） ， -
stimulation doses for subsequent experiments. ii Compared with the control group the activities of SOD and GSH Px in L02
（ P ） （ P ）， -
cells in the three doses groups decreased all <0.05 and the levels of MAD increased all <0.05 with a dose effect
- （P ），
relationship. The CAT activity of L02 cells in the medium dose group was lower than that in the control group <0.05 and the
- （ P ）
CAT activity of L02 cells in the high dose group was lower than that in the others three groups all <0.05 . Compared with the
， NRF2 - （P ），NRF2
control group the relative expression levels of mRNA in L02 cells in the low dose group decreased <0.05
- （P ）， NRF2
mRNA in L02 cells in the medium dose group increased <0.05 mRNA and NRF2 protein expression in L02 cells in
（ P ） HO-1，GCLC， NQO1 ，
the highdose group increased both <0.05 . The relative expression level of mRNA and GCLC NQO1
（ P ）
protein expression in L02 cells in the three doses groups increased compared with the control group all <0.05 . The relative
NRF2 - - -
expression level of mRNA in L02 cells in the high dose group was higher than that in the low and medium dose groups
（ P ）， - （P ），
both <0.05 and the relative expression of NRF2 protein was higher than that in the low dose group <0.05 but the
HO-1 GCLC - - （
relative expression levels of and mRNA and HO 1 protein level were lower than those in the medium dose group all
P ）Conclusion -
<0.05 . TCM exposure can inhibit the proliferation of L02 cells by inducing oxidative stress with a dose effect
，
relationship. In this process the antioxidant mechanism mediated by NRF2 was activated. The expression of antioxidant defense
， - ，
and detoxification related target genes downstream of NRF2 signaling pathway was activated and the expression of HO 1
-
GCLC and NQO1 was up regulated to alleviate the oxidative damage caused by TCM.
