摘要
【目的】探讨在高钙诱导细胞氧化应激下,高表达衰老标记蛋白30(SMP30)对人晶状体上皮细胞(HLEC)株SRA01/04增殖活力及抗氧化能力的影响。【方法】实验分3组:SMP30高表达组(OE,实验组)、NCOE组(阴性对照组)及SRA01/04组(空白对照组)。用OE及NCOE慢病毒载体分别转染SRA01/04细胞。15 mmol/LCaCl2处理细胞24 h建立高钙诱导模型。BrdU法检测细胞增殖、超氧化物歧化酶(SOD)、氧化型谷胱甘肽(GSSG)/总谷胱甘肽(T-GSH)法检测细胞氧化指标。【结果】荧光显微镜下可见各组转染细胞有大量绿色荧光蛋白(GFP)表达,转染效率接近80%,提示SMP30高表达SRA01/04细胞模型构建成功。在高钙状态下,OE组细胞相对增殖活力(3.89±0.20)和SOD活力(U/mg,47.5±4.3)均高于NCOE组(2.82±0.34、30.6±4.2)及SRA01/04组(2.96±0.25、26.8±1.5),OE组GSSG/T-GSH比值(2.36±0.51)低于NCOE组(16.36±2.48)及SRA01/04组(20.12±2.54),上述差异均有统计学意义(n=3,P<0.05),NCOE组与SRA01/04组相比差异均无统计学意义(n=3,P>0.05)。【结论】高表达SMP30增加SRA01/04(HLEC)细胞增殖活力、SOD活力及下降GSSG/T-GSH水平,提示SMP30在一定程度上可缓解高钙诱导细胞氧化损伤的进程,可能对HLEC有保护作用。
【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.
作者
李松蔓
陈曦
韩子豪
THU THU WIN AINT
衷昕
唐东永
梁皓
LI Song-man;CHEN Xi;HAN Zi-hao;THU THU WIN AINT;ZHONG Xin;TANG Dong-yong;LIANG Hao(Department of Ophthalmology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)
出处
《中山大学学报（医学版）》
CAS
CSCD
北大核心
2019年第1期54-61,共8页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金地区科学基金(81360146)
关键词
衰老标记蛋白30
人晶状体上皮细胞
高钙
氧化应激
超氧化物歧化酶
谷胱甘肽
细胞增殖
senescence marker protein 30
human lens epithelial cell
high calcium
oxidative stress
superoxide dismutase
glutathione
cell proliferation
