高级检索
中华实验眼科杂志
期刊首页
过刊列表
高级检索
稿件发表
• 实验研究 •
ENGLISH ABSTRACT
糖皮质激素对晶状体上皮细胞生物学功能调控作用的生物信息学分析
闫楚凡
韩笑
张劲松
作者及单位信息
·
DOI： 10.3760/cma.j.cn115989-20190821-00359
Regulatory effect of glucocorticoid on the biological function of lens epithelial cells: a bioinformatics analysis
Yan Chufan
Han Xiao
Zhang Jinsong
Authors Info & Affiliations
Yan Chufan
Department of Ophthalmology, The Fourth Affiliated Hospital of China Medical University, Shenyang 110059, China
Han Xiao
Department of Ophthalmology, The Fourth Affiliated Hospital of China Medical University, Shenyang 110059, China
Zhang Jinsong
Department of Ophthalmology, The Fourth Affiliated Hospital of China Medical University, Shenyang 110059, China
·
DOI： 10.3760/cma.j.cn115989-20190821-00359
536
34
0
0
1
0
PDF下载
APP内阅读
摘要

目的利用生物信息学方法分析糖皮质激素对晶状体上皮细胞(LECs)生物学功能的影响，并预测相关的微小RNA(miRNA)。

方法下载GSE3040数据集，设置1 μmol/L地塞米松处理的人LECs细胞株HLE-B3细胞为实验组，1 μmol/L二甲基亚砜(DMSO)处理的HLE-B3细胞为对照组，利用GEO2R工具分析2个组间的差异表达基因。利用Metascape网站进行差异基因功能富集分析，通过EdU细胞增生实验检测2个组间细胞增生差异。通过STRING网站和cytoscape软件构建蛋白互作网络图，cytohubba app计算出hub基因，采用实时荧光定量PCR法检测2个组间hub基因的表达差异。利用mirCode数据库预测与hub基因相关的miRNA。

结果实验组与对照组间分析出341个差异基因，其中上调基因为300个，下调基因为41个。下调基因中差异最显著的5个基因是 SLC12A1、 MED13L、 ALDH5A1、 SLC15A3和 WWC1基因；上调基因中差异最显著的5个基因是 SCNN1A、 ANKRD36、 FKBP5、 PYY和 ADH1B基因。列出了富集量排名前20的生物学功能关键词，结果显示富集量最大的是对HLE-B3细胞增生的负向调节。实验组细胞增生率为(8.09±0.20)%，低于对照组的(39.63±0.80)%，差异有统计学意义( t＝38.43， P<0.01)。前10位hub基因分别是 SST、 CXCL8、 GRM1、 GNRH1、 CXCL5、 PPBP、 CX3CR1、 PYY、 EDNRA和 GRK5，实时荧光定量PCR结果显示2个组间SST、CXCL8、GRM1、PYY、EDNRA和GRK5 mRNA相对表达量比较差异均有统计学意义(均 P<0.05)。与hub基因相关性强的前6位miRNA分别是miR-15abc、miR-214、miR-23abc、miR-129-5p、miR-132和miR-24。

结论1 μmol/L地塞米松即可对HLE-B3细胞的增生产生负性调控作用。 SST、 CXCL8、 GRM1、 PYY、 EDNRA和 GRK5基因可能是糖皮质激素的作用靶点，miR-15abc、miR-214、miR-23abc、miR-129-5p、miR-132和miR-24最可能与hub基因相关。

白内障;糖皮质激素;生物信息学分析;晶状体上皮细胞;生物学功能;Hub基因;微小RNA
ABSTRACT

ObjectiveTo analyze the effect of glucocorticoid on the biological function of lens epithelial cells (LECs) by bioinformatics and predict related microRNA (miRNA).

MethodsGSE3040 database was downloaded and the human LECs line (HLE-B3) cells in the experimental group were treated with 1 μmol/L dexamethasone, and HLE-B3 cells in the control group were treated with 1 μmol/L dimethyl sulfoxide(DMSO).GEO2R was used to analyze the differentially expressed genes between the two groups.Metascape website was employed to analyze the functional enrichment of differentially expressed genes, and EdU cell proliferation assay was performed to detect the difference in cell proliferation between the two groups.STRING website and cytoscape software were used to construct protein-protein interaction network.Hub genes were calculated by cytohubba app, and quantitative real-time PCR was performed to detect the expression levels of hub genes between the two groups.MirCode website was used to predict the related miRNAs.

ResultsA total of 341 differentially expressed genes were detected between the experimental group and the control group, among which there were 300 up-regulated genes and 41 down-regulated genes. SLC12A1, MED13L, ALDH5A1, SLC15A3 and WWC1 were the top five down-regulated genes and SCNN1A, ANKRD36, FKBP5, PYY and ADH1B were the top five up-regulated genes.The top 20 terms of functional enrichment were listed, and the negative regulation of HLE-B3 cells proliferation showed the most enrichment.Cell proliferation rate in the experimental group was (8.09±0.20)%, which was significantly lower than (39.63±0.80)% in the control group ( t=38.43, P<0.01).The top ten hub genes were SST, CXCL8, GRM1, GNRH1, CXCL5, PPBP, CX3CR1, PYY, EDNRA and GRK5, and quantitative real time PCR confirmed that the expression levels of SST, CXCL8, GRM1, PYY, EDNRA and GRK5 mRNA were statistically different (all at P<0.05).The top six miRNAs which might be associated with hub genes were miR-15abc, miR-214, miR-23abc, miR-129-5p, miR-132 and miR-24.

ConclusionsThe 1 μmol/L glucocorticoid can negatively regulate the proliferation of HLE-B3 cells. SST, CXCL8, GRM1, PYY, EDNRA and GRK5 may be hub genes and miR-15abc, miR-214, miR-23abc, miR-129-5p, miR-132, miR-24 are most likely to relate to them.

Cataract;Glycocorticoid;Bioinformatics analysis;Lens epithelial cells;Biological function;Hub gene;MicroRNA
Zhang Jinsong, Email: mocdef.3ab61sjz_h4umc
Copyright by Chinese Medical Association
No content published by the journals of Chinese Medical Association may be reproduced or abridged without authorization. Please do not use or copy the layout and design of the journals without permission.
All articles published represent the opinions of the authors, and do not reflect the official policy of the Chinese Medical Association or the Editorial Board, unless this is clearly specified.
引用本文

闫楚凡,韩笑,张劲松. 糖皮质激素对晶状体上皮细胞生物学功能调控作用的生物信息学分析[J]. 中华实验眼科杂志,2021,39(04)：297-303.

DOI:10.3760/cma.j.cn115989-20190821-00359

PERMISSIONS

Request permissions for this article from CCC.

评价本文
*以上评分为匿名评价

版权归中华医学会所有。

未经授权，不得转载、摘编本刊文章，不得使用本刊的版式设计。

除非特别声明，本刊刊出的所有文章不代表中华医学会和本刊编委会的观点。

糖皮质激素性白内障多见于全身或局部长期使用糖皮质激素者，其发病机制尚未完全阐明。许多生物学过程被证明与糖皮质激素性白内障的发生相关，如细胞分化异常和糖皮质激素受体(glucocorticoid receptor，GR)的激活等 [ 1 , 2 , 3 ]；糖皮质激素还导致晶状体中E钙黏附蛋白和N钙黏附蛋白的表达量下降，而细胞黏附在晶状体上皮细胞(lens epithelial cells，LECs)的正常分化和迁移中有重要作用 [ 4 ]。微小RNA(microRNA，miRNA)是一种长度为18～22个核苷酸的非编码RNA，与基因的3’端非翻译区结合，通过抑制基因的转录来影响蛋白的表达。既往研究发现，miR-29a可以靶向抑制GR mRNA的表达，同时其本身的表达量又依赖于GR的激活，形成一个负反馈回路，这个特点在糖皮质激素治疗过程中有助于减缓GR水平的逐渐降低 [ 5 ]，这为糖皮质激素性白内障的治疗提供了新的思路。目前关于miRNA与糖皮质激素性白内障的研究尚未见报道。本研究基于生物信息学方法分析经地塞米松处理的LECs中差异基因的表达及其相关的生物学功能，筛选并验证hub基因的表达，预测与其最有可能相关的miRNA，以期为白内障的治疗提供参考。
试读结束，您可以通过登录机构账户或个人账户后获取全文阅读权限。
参考文献
[1]
Wang C , Dawes LJ , Liu Y ，et al.Dexamethasone influences FGF-induced responses in lens epithelial explants and promotes the posterior capsule coverage that is a feature of glucocorticoid-induced cataract[J ]. Exp Eye Res， 2013，111:79-87. DOI: 10.1016/j.exer.2013.03.006 .
返回引文位置
[2]
Gupta V , Wagner BJ .Expression of the functional glucocorticoid receptor in mouse and human lens epithelial cells[J ]. Invest Ophthalmol Vis Sci， 2003，44(5):2041-2046. DOI: 10.1167/iovs.02-1091 .
返回引文位置
[3]
王祥群，吴开力，曾骏文．糖皮质激素诱发白内障与Ca介导效应的实验研究[J]. 中华实验眼科杂志， 2005，23(2):125-127. DOI: 10.3760/cma.j.issn.2095-0160.2005.02.004 .
返回引文位置
Wang XQ , Wu KL , Zeng JW .Effects of calcium on glucocorticoid-induced cataract in chicken embryo[J ]. Chin J Exp Ophthalmol， 2005，23(2):125-127. DOI: 10.3760/cma.j.issn.2095-0160.2005.02.004 .
Goto Citation
[4]
Lyu J , Kim JA , Chung SK ，et al.Alteration of cadherin in dexamethasone-induced cataract organ-cultured rat lens[J ]. Invest Ophthalmol Vis Sci， 2003，44(5):2034-2040. DOI: 10.1167/iovs.02-0602 .
返回引文位置
[5]
Glantschnig C , Koenen M , Gil-Lozano M ，et al.A miR-29a-driven negative feedback loop regulates peripheral glucocorticoid receptor signaling[J ]. FASEB J， 2019，33(5):5924-5941. DOI: 10.1096/fj.201801385RR .
返回引文位置
[6]
Gupta V , Galante A , Soteropoulos P ，et al.Global gene profiling reveals novel glucocorticoid induced changes in gene expression of human lens epithelial cells[J ]. Mol Vis， 2005，11:1018-1040.
返回引文位置
[7]
Barrett T , Wilhite SE , Ledoux P ，et al.NCBI GEO：archive for functional genomics data sets—update[J/OL ]. Nucleic Acids Res， 2013，41(Database issue):D991-995[2020-06-23]. https://pubmed.ncbi.nlm.nih.gov/23193258/. DOI: 10.1093/nar/gks1193 .
返回引文位置
[8]
Zhou Y , Zhou B , Pache L ，et al.Metascape provides a biologist-oriented resource for the analysis of systems-level datasets[J/OL ]. Nat Commun， 2019，10(1):1523[2020-06-25]. https://pubmed.ncbi.nlm.nih.gov/30944313/. DOI: 10.1038/s41467-019-09234-6 .
返回引文位置
[9]
Smoot ME , Ono K , Ruscheinski J ，et al.Cytoscape 2.8：new features for data integration and network visualization[J ]. Bioinformatics， 2011，27(3):431-432. DOI: 10.1093/bioinformatics/btq675 .
返回引文位置
[10]
Jeggari A , Marks DS , Larsson E ．miRcode：a map of putative microRNA target sites in the long non-coding transcriptome[J]. Bioinformatics， 2012，28(15):2062-2063. DOI: 10.1093/bioinformatics/bts344 .
返回引文位置
[11]
Black RL , Oglesby RB , von Sallmann L ，et al.Posterior subcapsular cataracts induced by corticosteroids in patients with rheumatoid arthritis[J ]. JAMA， 1960，174:166-171. DOI: 10.1001/jama.1960.63030020005014 .
返回引文位置
[12]
Ernst P , Baltzan M , Deschênes J ，et al.Low-dose inhaled and nasal corticosteroid use and the risk of cataracts[J ]. Eur Respir J， 2006，27(6):1168-1174. DOI: 10.1183/09031936.06.00043005 .
返回引文位置
[13]
Sulaiman RS , Kadmiel M , Cidlowski JA .Glucocorticoid receptor signaling in the eye[J ]. Steroids， 2018，133:60-66. DOI: 10.1016/j.steroids.2017.11.002 .
返回引文位置
[14]
Carrasco E , Hernández C , Miralles A ，et al.Lower somatostatin expression is an early event in diabetic retinopathy and is associated with retinal neurodegeneration[J ]. Diabetes Care， 2007，30(11):2902-2908. DOI: 10.2337/dc07-0332 .
返回引文位置
[15]
Simó R , Carrasco E , Fonollosa A ，et al.Deficit of somatostatin in the vitreous fluid of patients with diabetic macular edema[J ]. Diabetes Care， 2007，30(3):725-727. DOI: 10.2337/dc06-1345 .
返回引文位置
[16]
Cheung CM , Vania M , Ang M ，et al.Comparison of aqueous humor cytokine and chemokine levels in diabetic patients with and without retinopathy[J ]. Mol Vis， 2012，18:830-837.
返回引文位置
[17]
Na KS , Mok JW , Kim JY ，et al.Correlations between tear cytokines，chemokines，and soluble receptors and clinical severity of dry eye disease[J ]. Invest Ophthalmol Vis Sci， 2012，53(9):5443-5450. DOI: 10.1167/iovs.11-9417 .
返回引文位置
[18]
Vishnivetskiy SA , Ostermaier MK , Singhal A ，et al.Constitutively active rhodopsin mutants causing night blindness are effectively phosphorylated by GRKs but differ in arrestin-1 binding[J ]. Cell Signal， 2013，25(11):2155-2162. DOI: 10.1016/j.cellsig.2013.07.009 .
返回引文位置
[19]
Ehlken C , Grundel B , Michels D ，et al.Increased expression of angiogenic and inflammatory proteins in the vitreous of patients with ischemic central retinal vein occlusion[J/OL ]. PLoS One， 2015，10(5):e0126859[2020-06-27]. https://pubmed.ncbi.nlm.nih.gov/25978399/. DOI: 10.1371/journal.pone.0126859 .
返回引文位置
[20]
Roche SL , Wyse-Jackson AC , Ruiz-Lopez AM ，et al.Fractalkine-CX3CR1 signaling is critical for progesterone-mediated neuroprotection in the retina[J/OL ]. Sci Rep， 2017，7:43067[2020-06-28]. https://pubmed.ncbi.nlm.nih.gov/28216676/. DOI: 10.1038/srep43067 .
返回引文位置
[21]
Zabel MK , Zhao L , Zhang Y ，et al.Microglial phagocytosis and activation underlying photoreceptor degeneration is regulated by CX3CL1-CX3CR1 signaling in a mouse model of retinitis pigmentosa[J ]. Glia， 2016，64(9):1479-1491. DOI: 10.1002/glia.23016 .
返回引文位置
[22]
Patel C , Narayanan SP , Zhang W ，et al.Activation of the endothelin system mediates pathological angiogenesis during ischemic retinopathy[J ]. Am J Pathol， 2014，184(11):3040-3051. DOI: 10.1016/j.ajpath.2014.07.012 .
返回引文位置
[23]
Rupaimoole R , Slack FJ .MicroRNA therapeutics：towards a new era for the management of cancer and other diseases[J ]. Nat Rev Drug Discov， 2017，16(3):203-222. DOI: 10.1038/nrd.2016.246 .
返回引文位置
[24]
Cortez MA , Valdecanas D , Zhang X ，et al.Therapeutic delivery of miR-200c enhances radiosensitivity in lung cancer[J ]. Mol Ther， 2014，22(8):1494-1503. DOI: 10.1038/mt.2014.79 .
返回引文位置
[25]
Huang J , Chen Z , Lai Z ，et al.Kaempferol ameliorates the regulatory effects of PVT1/miR-214 on epithelial-mesenchymal transition through the PAK4/β-catenin axis in SRA01/04 cells[J/OL ]. Future Med Chem， 2021[2021-02-25]. https://pubmed.ncbi.nlm.nih.gov/33527844/. DOI:10.4155/fmc-2020-0381.[published online ahead of print Feb 2，2021].
返回引文位置
[26]
Yang J , Zhao S , Tian F ．SP1-mediated lncRNA PVT1 modulates the proliferation and apoptosis of lens epithelial cells in diabetic cataract via miR-214-3p/MMP2 axis[J]. J Cell Mol Med， 2020，24(1):554-561. DOI: 10.1111/jcmm.14762 .
返回引文位置
[27]
Wu C , Lin H , Wang Q ，et al.Discrepant expression of microRNAs in transparent and cataractous human lenses[J ]. Invest Ophthalmol Vis Sci， 2012，53(7):3906-3912. DOI: 10.1167/iovs.11-9178 .
返回引文位置
[28]
Lu B , Christensen IT , Ma LW ，et al.miR-24-p53 pathway evoked by oxidative stress promotes lens epithelial cell apoptosis in age-related cataracts[J ]. Mol Med Rep， 2018，17(4):5021-5028. DOI: 10.3892/mmr.2018.8492 .
返回引文位置
备注信息
A
张劲松，Email： mocdef.3ab61sjz_h4umc
B
所有作者均声明不存在任何利益冲突
C
国家自然科学基金项目 (81870644)
评论 （0条）
注册
登录
时间排序
暂无评论，发表第一条评论抢沙发
最新推荐
更多
丁型肝炎诊疗现状及药物研发进展
饶慧瑛 等
中华肝脏病杂志 2024,32(10)
中国甲状腺相关眼病诊断和治疗指南（2022年）
中华医学会眼科学分会眼整形眼眶病学组 等
中华眼科杂志 2022,58(09)
脓毒症基础研究的进展及未来方向
张笑婷 等
中华危重病急救医学 2021,33(08)
重视后发性白内障防治的应用基础研究
汤欣 等
中华实验眼科杂志 2018,36(03)
MedAI助手（体验版）
文档即答
智问智答
机器翻译
回答内容由人工智能生成，我社无法保证其准确性和完整性，该生成内容不代表我们的态度或观点，仅供参考。
生成快照
文献快照

你好，我可以帮助您更好的了解本文，请向我提问您关注的问题。

0/2000

《中华医学会杂志社用户协议》 | 《隐私政策》

《SparkDesk 用户协议》 | 《SparkDesk 隐私政策》

网信算备340104764864601230055号 | 网信算备340104726288401230013号

技术支持：

历史对话
本文全部
还没有聊天记录
设置
模式
纯净模式沉浸模式
字号
小中大