[1]李兰根,伟伟,张玉凤,等.视网膜色素上皮细胞氧化应激过程中SirT1/STAT3的相互作用[J].眼科新进展,2017,37(12):1119-1122.[doi:10.13389/j.cnki.rao.2017.0282]
　LI Lan-Gen,WEI Wei,ZHANG Yu-Feng,et al.Interactions between SirT1 and STAT3 in the retinal pigmented epithelium cell[J].Recent Advances in Ophthalmology,2017,37(12):1119-1122.[doi:10.13389/j.cnki.rao.2017.0282]
点击复制

视网膜色素上皮细胞氧化应激过程中SirT1/STAT3的相互作用/HTML
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
37卷
期数:
2017年12期
页码:
1119-1122
栏目:
实验研究
出版日期:
2017-12-05

文章信息/Info

Title:
Interactions between SirT1 and STAT3 in the retinal pigmented epithelium cell
作者:
李兰根; 伟伟; 张玉凤; 格日乐图; 张艳梅
010017　内蒙古自治区呼和浩特市，内蒙古自治区人民医院眼科（李兰根，伟伟，张玉凤，格日乐图）；010017　内蒙古自治区呼和浩特市，内蒙古自治区人民医院神经内科（张艳梅）
Author(s):
LI Lan-Gen; WEI Wei; ZHANG Yu-Feng; GE-RI Le-Tu; ZHANG Yan-Mei
Departments of Ophthalmology，Inner Mongolia People’s Hospital（LI Lan-Gen，WEI Wei，ZHANG Yu-Feng，GE-RI Le-Tu），Hohhot 010017，
关键词:
去乙酰化酶-1; 信号转导和转录激活因子3; 视网膜色素上皮细胞; 细胞氧化应激
Keywords:
SirT1; STAT3; retinal pigment epithelial cells; oxidative stress
分类号:
R774
DOI:
10.13389/j.cnki.rao.2017.0282
文献标志码:
A
摘要:
目的　探讨去乙酰化酶-1（silent information regulator of transcription 1，SirT1）和信号转导和转录激活因子3（signal transducer and activator of transcription 3，STAT3）在视网膜色素上皮（retinal pigment epithelium，RPE）细胞氧化应激过程中的相互作用。方法　体外培养RPE细胞系ARPE-19，H2O2诱导氧化应激。分别采用靶向敲除策略（siRNA/shRNA）沉默SirT1/STAT3基因及采用慢病毒载体（pRC/CMV STAT3及pRC/CMV）转染宿主细胞诱导过表达；采用 RT-PCR及Western blot观察基因敲除/过表达状态下SirT1/STAT3表达的相互影响及SirT1/STAT3相互作用对暴露于氧化应激状况下RPE细胞的影响。结果　SirT1激活剂白藜芦醇能显著降低STAT3 mRNA的表达量3.0±0.2（P=0.048）；而SirT1基因敲除后，RPE细胞中STAT3 mRNA表达量（6.9±1.1）显著上升（P=0.025）；在SirT1基因敲除后RPE细胞氧化应激过程中STAT3蛋白和磷酸化STAT3蛋白表达显著提高，分别为0.990±0.031、0.544±0.019（P=0.000、0.003）。氧化应激条件下STAT3过表达能显著诱发SirT1 mRNA表达（0.42±0.16）水平下降（P=0.022）；而STAT3敲除后SirT1 mRNA表达量（2.80±0.85）显著增加（P=0.015）。结论　RPE细胞氧化应激过程中SirT1对STAT3表达起到负向调节作用，提示在RPE氧化应激过程中SirT1和STAT3之间存在平衡机制。
Abstract:
Objective　To investigate the interactions between histone deacetylas-1 （silent information regulator of transcription 1，SirT1） and signal transducer and activator of transcription 3 （STAT3） following exposure to oxidative stress of retinal pigmented epithelium （RPE） cells through gene knockout and overexpression techniques.Methods　RPE cell line was cultured in vitro，followed by addition of H2O2 for inducing oxidative stress.The targeting knockout strategy （siRNA/shRNA） was used for silencing SirT1/STAT3 gene and lentiviral vectors （pRC/CMV STAT3 and PCRC/CMV） were transfected to RPE cells.RT-qPCR and Western blot were used to detect the expression of SirT1/STAT3 due to gene knockout and overexpression and deeply analyze the effects of SirT1/STAT3 on RPE cells exposed to oxidative stress.Results　SirT1 activator （resveratrol） significantly reduced the expression of STAT3 mRNA to （3.0±0.2） （P=0.048）；and after SirT1 knockout，the expression of STAT3 mRNA in RPE cells was significantly increased to （6.9±1.1） （P=0.025）.During the oxidative stress of RPE cells after SirT1 knockout，the expression of STAT3 and phosphorylated STAT3 protein were significantly increased to 0.990±0.031 and 0.544±0.019，respectively （P=0.000，0.003）.With the condition of oxidative stress，the over-expression of STAT3 reduced the SirT1 mRNA expression to 0.42±0.16 （P=0.022），but SirT1 mRNA expression was significantly increased to 2.8±0.85 （P=0.015） during STAT3 knockdown.Conclusion　SirT1 has a negative effect on the regulation of STAT3 expression during oxidative stress，which suggests that there is an equilibrium mechanism between SirT1 and STAT3 against oxidative stress.

参考文献/References:

［1］　LI Z，DONG X，LIU H，CHEN X，SHI H，FAN Y，et al.Astaxanthin protects ARPE 19 cells from oxidative stress via upregulation of Nrf2 regulated phase II enzymes through activation of PI3K/Akt［J］.Mol Vis，2013，19：1656-1666.
［2］　GORIN MB.A new vision for age-related macular degeneration［J］.Eur Hun Genet，2005，13（7）：793-794.
［3］　WEI W，LANGEN L，YUFENG Z，GERILETU，JIA Y，YANMEI Z.Vitamin C protected human retinal pigmented epithelium from oxidant injury depending on regulating SIRT1［J］.Scientific World J，2014，2014：750634.
［4］　KAGAN DB，LIU H，HUTNIK CM.Efficacy of various antioxidants in the protection of the retinal pigment epithlium from oxidative stress［J］.Clin Opthalmol，2012，6：1471-1476.
［5］　TANNO M，KUNO A，YANO T，MIURA T，HISAHARA S，ISHIKAWA S，et al.Induction of manganese superoxide dismutase by nuclear translocation and activation of SIRT1 promotes cell survival in chronic heart failure［J］.Biol Chem，2010，285（11）：8375-8382.
［6］　VINCIGUERRA M，SANTINI MP，MARTNEZ C，PAZIENZA V，CLAYCOMB WC，GIULIANI A，et al.mIGF 1/JNK1/SirT1 signaling confers protection against oxidative stress in the heart［J］.Aging Cell，2012，11（1）：139-149.
［7］　CAITO S，RAJENDRASOZHAN S，COOK S.SIRT1 is a redox sensitive deacetylase that is post translationally modified by oxidants and carbonyl stress［J］.FASEB，2010，24（9）：3145-3159.
［8］　KAWAI Y，GARDUNO L，THEODORE M，YANG J ARINZE IJ.Acety lation deacetylation of the transcription factor Nrf2 （nuclear factor erythroid 2 related factor 2） regulates its transcriptional activity and nucleocytoplasmic localization［J］.Biol Chem，2011，286（9）：7629-7640.
［9］　PATEL AK，HACKAM AS.Toll like receptor 3 （TLR3） protects retinal pigmented epithelium （RPE） cells from oxidative stress through a STAT3 dependent mechanism［J］.Mol Immunol，2013，54（2）：122-131.
［10］　BERNIER M，PAUL RK，MARTIN MONTALVO A，Scheibye-Knudsen M，Song S，He HJ，et al.Negative regulation of STAT3 protein mediated cellular respiration by SIRT1 protein［J］.Biol Chem，2011，286（22）：19270-19279.
［11］　JARRETT SG，BOULTON ME.Consequences of oxidative stress in age related macular degeneration［J］.Mol Aspects Med，2012，33（4）：399-417.
［12］　ZHANG C，LI H，LIU MG，Kawasaki A，Fu XY，Barnstable CJ，et al.STAT3 activation protects retinal ganglion cell layer neurons in response to stress［J］.Exp Eye Res，2008，86（6）：991-997.
［13］　MIMURA T，KAJI Y，NOMA H，FUNATSU H，OKAMOTO S.The role of SIRT1 in ocular aging［J］.Exp Eye Res，2013，116：17-26.
［14］　PLAFKER SM，O’Mealey GB，SZWEDA LI.Mechanisms for counterin oxidative stress and damage in retinal pigment epithelium［J］.Int Rev Cell Mol Biol，2012，298：135-177.

备注/Memo

备注/Memo:
内蒙古自治区自然科学基金资助项目（编号：2016MS0872）
更新日期/Last Update: 2017-12-08