[1]季青山,俞茜,孙思勤,等.miR-34a/SIRT1在H2O2诱导人晶状体上皮细胞氧化应激中的表达[J].眼科新进展,2017,37(8):728-731.[doi:10.13389/j.cnki.rao.2017.0184]
 JI Qing-Shan,YU Xi,SUN Si-Qin,et al.Expression of miR-34a/SIRT1 in human lens epithelial cells during H2O2-induced oxidative stress[J].Recent Advances in Ophthalmology,2017,37(8):728-731.[doi:10.13389/j.cnki.rao.2017.0184]
点击复制

miR-34a/SIRT1在H2O2诱导人晶状体上皮细胞氧化应激中的表达/HTML
分享到:

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年8期
页码:
728-731
栏目:
实验研究
出版日期:
2017-08-05

文章信息/Info

Title:
Expression of miR-34a/SIRT1 in human lens epithelial cells during H2O2-induced oxidative stress
作者:
季青山俞茜孙思勤柯根杰温跃春
230001 安徽省合肥市,安徽医科大学附属省立医院眼科
Author(s):
JI Qing-ShanYU XiSUN Si-QinKE Gen-JieWEN Yue-Chun
Department of Ophthalmology,the Affiliated Provincial Hospital,Anhui Medical University,Hefei 230001,Anhui Province,China
关键词:
微小RNA-34a沉默信息调节因子1过氧化氢人晶状体上皮细胞氧化应激
Keywords:
microRNA-34asilent information regulator 1hydrogen peroxidehuman lens epithelial celloxidative stress
分类号:
R776.1
DOI:
10.13389/j.cnki.rao.2017.0184
文献标志码:
A
摘要:
目的 观察微小RNA-34a(microRNA-34a,miR-34a)和沉默信息调节因子1(silent information regulator 1,SIRT1)在不同浓度 H2O2诱导的人晶状体上皮细胞(SRA01/04细胞)氧化应激中的表达变化。方法 用不同浓度的 H2O2(0 μmol·L-1、100 μmol·L-1、200 μmol·L-1、300 μmol·L-1、400 μmol·L-1)处理细胞24 h后,用CCK-8检测细胞存活率,流式细胞仪检测细胞凋亡率和RT-PCR检测miR-34a/SIRT1的表达。结果 CCK-8检测结果显示:100~400 μmol·L-1 H2O2对SRA01/04细胞有增殖抑制作用,且呈剂量依赖关系,与0 μmol·L-1 H2O2组相比差异均有统计学意义(均为P<0.01);流式细胞仪凋亡检测结果显示:0 μmol·L-1 H2O2组细胞凋亡率为(6.1±1.2)%;100 μmol·L-1、200 μmol·L-1、300 μmol·L-1 H2O2组细胞凋亡率分别为(26.3±1.8)%、(32.5±2.2)%、(64.7±5.3)%,各组与0 μmol·L-1 H2O2组比较,差异均有统计学意义(均为P<0.01)。RT-PCR检测结果显示:不同浓度 H2O2处理SRA01/04细胞后,细胞中的miR-34a表达水平呈剂量依赖性升高,而SIRT1表达水平呈相应下降,与0 μmol·L-1 H2O2组相比差异均有统计学意义(均为P<0.001)。结论 在一定浓度 H2O2诱导的人晶状体上皮细胞氧化应激中,miR-34a表达水平显著增加,而SIRT1表达水平显著下降。下调miR-34a表达可增加氧化应激人晶状体上皮细胞存活率。
Abstract:
Objective To investigate the expression of microRNA-34a (miR-34a) and silent information regulator 1 (SIRT1) in human lens epithelial cells under H2O2-induced oxidative stress.Methods Different concentrations of H2O2 (0 μmol·L-1,100 μmol·L-1,200 μmol·L-1,300 μmol·L-1,and 400 μmol·L-1) were used to stimulate SRA01/04 cells for 24 hours.Cell viability was measured using cell counting kit-8 (CCK-8) assay.Cell apoptosis was detected by flow cytometry.Expression levels of miR-34a/SIRT1 were measured by RT- PCR.Results CCK-8 assay showed that a certain concentration range of H2O2 had a proliferation inhibition on SRA01/04 cells.There was a dose response relationship between 100 μmol·L-1 and 400 μmol·L-1.Compared with 0 μmol·L-1 H2O2 group,the difference was statistically significant (all P<0.01).According to flow cytometry results,apoptotic rate of SRA01/04 cells in control group and H2O2 (100-300 μmol·L-1) groups were (6.1±1.2)%,(26.3±1.8)%,(32.5±2.2)%,and (64.7±5.3)%.Compared with 0 μmol·L-1 H2O2 group,the differences were statistically significant (all P<0.01).RT-PCR test results showed that the expression of miR-34a increased significantly in a dose-dependent manner after the SRA01/04 cells treated with different concentrations of H2O2,while SIRT1 expression level was decreased,there were significant differences compared with control group (all P<0.001).Conclusion There is a significantly increase of miR-34a and decrease of SIRT1 in human lens epithelial cells under the oxidative stress of a certain concentration of H2O2.Down-regulated expression of miR-34a can increase the survival rate of human lens epithelial cells under H2O2-induced oxidative stress.

参考文献/References:

[1] SPECTOR A,GARNER WH.Hydrogen peroxide and human cataract[J].Exp Eye Res,1981,33(6):673-681.
[2] LI WC,KUSZAK JR,DUNN K,WANG RR,MA W,WANG GM,et al.Lens epithelial cell apoptosis appears to be a common cellular basis for non-congenital cataract development in humans and animals[J].J Cell Biol,1995,130(1):169-181.
[3] DUAN K,GE YC,ZHANG XP,WU SY,FENG JS,CHEN SL,et al.miR-34a inhibits cell proliferation in prostate cancer by downregulation of SIRT1 expression[J].Oncol Lett,2015,10(5):3223-3227.
[4] SAITO Y,NAKAOKA T,SAITO H.microRNA-34a as a Therapeutic Agent against Human Cancer[J].J Clin Med,2015,4(11):1951-1959.
[5] LU G,SUN Y,AN S,XIN S,REN X,ZHANG D,et al.MicroRNA-34a targets FMNL2 and E2F5 and suppresses the progression of colorectal cancer[J].Exp Mol Pathol,2015,99(1):173-179.
[6] CHIEN KH,CHEN SJ,LIU JH,CHANG HM,WOUNG LC,LIANG CM,et al.Correlation between microRNA-34a levels and lens opacity severity in age-related cataracts[J].Eye,2013,27(7):883-888.
[7] YANG Y,SHARMA R,CHENG JZ,SAINI MK,ANSARI NH,ANDLEY UP,et al.Protection of HLE B-3 cells against hydrogen peroxide-and naphthalene-induced lipid peroxidation and apoptosis by transfection with hGSTA1 and hGSTA2[J].Invest Ophthalmol Vis Sci,2002,43(2):434-445.
[8] PENG Y,GUO JJ,LIU YM,WU XL.MicroRNA-34A inhibits the growth,invasion and metastasis of gastric cancer by targeting PDGFR and MET expression[J].Biosci Rep,2014,34(3).pii:e00112.
[9] SCHIRMER U,DOBERSTEIN K,RUPP AK,BRETZ NP,WUTTIG D,KIEFEL H,et al.Role of miR-34a as a suppressor of L1CAM in endometrial carcinoma[J].Oncotarget,2014,5(2):462-472.
[10] STASZEL T,ZAPALA B,POLUS A,SADAKIERSKA CA,KIEC WB,STEPIEN E,et al.Role of microRNAs in endothelial cell pathophysiology[J].Pol Arch Med Wewn,2011,121(10):361-366.
[11] ZHU L,GAO J,HUANG K,LUO Y,ZHANG B,XU W.miR-34a screened by miRNA profiling negatively regulates Wnt/beta-catenin signaling pathway in Aflatoxin B1 induced hepatotoxicity[J].Sci Rep,2015,5:16732.
[12] PENG CH,CHANG YL,KAO CL,TSENG LM,WU CC,CHEN YC,et al.SirT1-a sensor for monitoring self-renewal and aging process in retinal stem cells[J].Sensors(Basel),2010,10(6):6172-6194.
[13] TABUCHI T,SATOH M,ITOH T,NAKAMUR A.MicroRNA-34a regulates the longevity-associated protein SIRT1 in coronary artery disease:effect of statins on SIRT1 and microRNA-34a expression[J].Clin Sci(Lond),2012,123(3):161-171.
[14] XIONG H,PANG J,YANG H,DAI M,LIU Y,OU Y,et al.Activation of miR-34a/SIRT1/p53 signaling contributes to cochlear hair cell apoptosis:implications for age-related hearing loss[J].Neurobiol Aging,2015,36(4):1692-1701.

相似文献/References:

[1]康丽华,邹茜,杨梅,等.年龄相关性白内障患者沉默信息调节因子1(SIRT1)的单核苷酸多态性研究[J].眼科新进展,2017,37(10):939.[doi:10.13389/j.cnki.rao.2017.0238]
 KANG Li-Hua,ZOU Xi,YANG Mei,et al.Association of single nucleotide polymorphism of SIRT1 gene with age-related cataract[J].Recent Advances in Ophthalmology,2017,37(8):939.[doi:10.13389/j.cnki.rao.2017.0238]
[2]陆博,马立威,王欣玲,等.年龄相关性白内障患者miR-138的表达及其对人晶状体上皮细胞凋亡的影响[J].眼科新进展,2018,38(2):111.[doi:10.13389/j.cnki.rao.2018.0024]
 LU Bo,MA Li-Wei,WANG Xin-Ling,et al.Expression of miR-138 and its effects on human lens epithelial cells apoptosis in age-related cataract patients[J].Recent Advances in Ophthalmology,2018,38(8):111.[doi:10.13389/j.cnki.rao.2018.0024]
[3]潘家钰,康刚劲,徐曼华,等.雌二醇通过激活SIRT1/P53通路对人晶状体上皮细胞抗凋亡作用研究[J].眼科新进展,2020,40(5):420.[doi:10.13389/j.cnki.rao.2020.0097]
 PAN Jiayu,KANG Gangjin,XU Manhua,et al.Anti-apoptotic effect of estradiol on human lens epithelial cells via SIRT1/P53 pathway[J].Recent Advances in Ophthalmology,2020,40(8):420.[doi:10.13389/j.cnki.rao.2020.0097]

备注/Memo

备注/Memo:
安徽省自然科学基金资助(编号:1508085SQH221)
更新日期/Last Update: 2017-08-14