Effect of Overexpressed Senescence Marker Protein 30 in Human Lens Epithelial Cell Line SRA01/04 under High-Calcium Mediated Oxidative Stress
- VernacularTitle:高表达衰老标记蛋白30对高钙诱导人晶状体上皮细胞株SRA01/04氧化应激的影响
- Author:
Song-man LI
1
;
Xi CHEN
1
;
Zi-hao HAN
1
;
THU THU WIN AINT
1
;
Xin ZHONG
1
;
Dong-yong TANG
1
;
Hao LIANG
1
Author Information
1. Department of Ophthalmology，the First Affiliated Hospital of Guangxi Medical University，Nanning 530021，China
- Publication Type:Journal Article
- Keywords:
senescence marker protein 30;
human lens epithelial cell;
high calcium;
oxidative stress;
superoxide dismutase;
glutathione;
cell proliferation
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2019;40(1):54-61
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】To explore the effects of overexpressed senescence marker protein 30 （SMP30） on cell proliferation and antioxidative activity in human lens epithelial cell（HLEC）line SRA01/04 under high-calcium mediated oxidative stress. 【Methods】There were 3 groups in this experiment：SMP30 overexpressed group （OE，experimental group），NCOE group （negative control group） and SRA01/04 group （blank control group）. OE and NCOE lentiviral vectors were used to transfect SRA01/04 respectively. A high- calcium- mediated- stress cell model was established by culturing cells with medium containing 15 mmol/L CaCl2 for 24 h. BrdU assay was used to measure cell proliferation. SOD assay kit and GSSG/T- GSH assay kit were used to detect the level of intracellular oxidative stress. 【Results】Green fluorescence protein could be observed in all transfected cell groups under fluorescence microscope and the transfection efficiency was close to 80% ，suggesting that OE cell model was constructed successfully. Under the high calcium culture conditions，the activity of relative cell proliferation and SOD in OE group［（3.89 ± 0.20）and（47.5 ± 4.3 U/mg）］were significantly higher than that in NCOE group［（2.82 ± 0.34）and（30.6 ± 4.2 U/mg）］and SRA01/04 group［（2.96 ± 0.25）and（26.8 ± 1.5 U/mg）］，the ratio of GSSG/T-GSH in OE group（2.36 ± 0.51）was significantly lower than that in NCOE group（16.36 ± 2.48）and SRA01/04 group（20.12 ± 2.54）（n=3，P<0.05）；there was no significant difference between NCOE group and SRA01/04 group （n=3，P>0.05）. 【Conclusions】Overexpression of SMP30 increased the activity of cell proliferation and SOD，but decreased the ratio of GSSG/T- GSH in SRA01/04 cell（HLEC），indicating that SMP30 may alleviate the progression of high-calcium-mediated oxidative cell damage and possess the cytoprotective functions in HLEC.
