[1]王嵩,梅林,闫博,等. 生理低氧对人晶状体上皮细胞系基因表达谱的影响[J].眼科新进展,2015,35(5):409-412.[doi:10.13389/j.cnki.rao.2015.0111]
　WANG Song,MEI Lin,YAN Bo,et al. Effects of physiological hypoxia on gene expression profile of human lens epithelial cells[J].Recent Advances in Ophthalmology,2015,35(5):409-412.[doi:10.13389/j.cnki.rao.2015.0111]
点击复制

 生理低氧对人晶状体上皮细胞系基因表达谱的影响
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年5期
页码:
409-412
栏目:
实验研究
出版日期:
2015-05-05

文章信息/Info

Title:
 Effects of physiological hypoxia on gene expression profile of human lens epithelial cells
作者:
 王嵩; 梅林; 闫博; 杨安钢; 赵晶; 严宏
 710038　陕西省西安市，第四军医大学唐都医院眼科
Author(s):
 WANG Song ; MEI Lin ;  YAN Bo ; YANG An-Gang; ZHAO Jing;  YAN Hong
 Department of Ophthalmology , Tangdu Hospital of the Fourth Military Medical University, Xi ’ an 7 10038 . Shaanxi Province , China
关键词:
 ; 人晶状体上皮细胞; 生理低氧; 基因表达谱
Keywords:
 ; human lens epithelial cells ;  physiological hypoxia;  gene expression profile
DOI:
10.13389/j.cnki.rao.2015.0111
文献标志码:
A
摘要:
 目的　探讨生理低氧条件下与常规氧环境下培养的人晶状体上皮细胞（ｈｕｍａｎｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｌ，ＨＬＥＣ）在抗氧化能力及基因表达谱上的差异。方法　ＨＬＥＣ传代培养２４ｈ后，分别置于体积分数２１％常氧培养箱、体积分数１％低氧培养箱孵育２４ｈ，消化、超速离心后收集细胞上清分别检测ＳＯＤ、ＣＡＴ活性以及ＧＳＨ含量，Ｔｒｉｚｏｌ提取细胞总ＲＮＡ进行基因表达谱检测，生物信息学软件ＤＡＶＩＤ分析差异基因的功能聚类，ｑＲＴ-ＰＣＲ验证差异基因。结果　低氧组细胞的ＳＯＤ、ＣＡＴ酶活性及ＧＳＨ含量均低于常氧组，差异均有统计学意义（均为Ｐ＜０．０５）。表达谱芯片结果显示：符合差异表达标准的基因中，在低氧组细胞上调的基因有３１９个，下调的基因有８３个。ＧｅｎｅＯｎｔｏｌｏｇｙ分析显示：３１９个上调基因聚类于１４８个功能分类中，富集值最高的功能分类为低氧反应。选取５个上调基因进行ｑＲＴ-ＰＣＲ验证显示：ＤＤＩＴ４、ＶＥＧＦＡ、ＥＤＮ１、ＶＬＤＬＲ、ＥＧＬＮ１在低氧组中的表达量均高于常氧组，差异均有统计学意义（均为Ｐ＜０．０５）。结论　生理低氧状态下ＨＬＥＣ的抗氧化能力弱于常氧组，两种氧环境下细胞的基因表达谱不同，提示相应的抗氧化损伤机制可能存在差异。
Abstract:
 Objective To compare the anti-oxidative capability and gene expression profile of human lens epithelial cells ( HLEC) cultured in both physiological hypoxia and normoxia. Methods After 24-hour subculture , HLEC were cultured in hypoxia and atmospheric oxygen respectively for another 24 hours. After trypsinized and centrifuged,HLEC supernatant was assayed for the amount of reduced glutathione ( GSH) and activities of superoxide dismutase ( SOD) and catalase ( CAT) . Total RNA was extracted by Trizol to detect gene expression profile. Bioinformatical database DAVID was used to perform gene ontology ( GO) analysis. The differentially expressed genes were validated by qRT-PCR. Results The amount of GSH and activities of SOD and CAT in the hypoxia group were lower than those in the normoxia group ( all P < 0. 05 ) . Gene expression profile indicated 319 up-regulated genes and 83 down-regulated genes in the hypoxia group. A1l the up-regulated genes were further enriched in 148 biological processes. Response to hypoxia was the most sigruficant process, among which 5 upregulated genes were further identified for their expression by qRT-PCR. It was shown that the expression level of DDIT4 , VEGFA, EDNI . VLDLR and EGLNI in the hypoxia group were higher than those in the normoxia group ( all P < 0. 05 ). Conclusion HLEC have weaker anti-oxidative ability in physiological hypoxia. Genes are differentially expressed in different oxygen environments ,which may contribute to differential antioxidative mechanisms.

相似文献/References:

[1]杨鑫 张凤妍 方梦园 张金嵩 彭广华.PUMA 介导氧化应激诱导的人晶状体上皮细胞凋亡[J].眼科新进展,2012,32(5):000.
[2]陈英 廖敏华 郑江娉 刘平.白藜芦醇对过氧化氢诱导人晶状体上皮细胞氧化损伤的保护作用[J].眼科新进展,2013,33(6):000.
[3]刘平 苏胜. 白内障蛋白质组学研究的现状及未来研究方向[J].眼科新进展,2014,34(1):001.
[4]杨宝娣 宋艳萍 陈中山 丁琴. 微脉冲半导体激光对兔视网膜色素上皮细胞阈值下光凝的光生物调制效应[J].眼科新进展,2014,34(1):005.
[5]赖伟霞 梁皓 谭少健 李霞 邹文进 蒋林志. 钙调素在正常人及不同年龄白内障患者晶状体上皮细胞中的表达[J].眼科新进展,2014,34(1):010.
[6]郑燕林 刘聪慧 沙菽. 水蛭提取液对体外培养RF/6A的细胞液中MMP-2、IV型胶原含量的影响[J].眼科新进展,2014,34(1):013.
[7]周琨 高晓唯 蔡岩 李文静 胡裕坤 田丽丽 付燕. CCR7基因修饰的未成熟树突状细胞在大鼠高危角膜移植免疫排斥反应中的作用[J].眼科新进展,2014,34(1):017.
[8]高伟 程燕 吴洁 王亮 赵帅. 肉毒杆菌B诱导大鼠干眼模型中泪腺Lacritin蛋白表达的研究[J].眼科新进展,2014,34(1):021.
[9]赵宁 张瑞君 刘磊 李佳 刘宁宁. PTEN基因对兔晶状体上皮细胞增殖抑制作用的实验研究[J].眼科新进展,2014,34(1):025.
[10]陈雪 李海平 张培 曹安民. 小胶质细胞在视网膜母细胞瘤中的活化及分布研究[J].眼科新进展,2014,34(1):029.
[11]杨楠,康刚劲,董敏. bFGF与ERK1/2阻断剂对人晶状体上皮细胞α平滑肌肌动蛋白mRNA表达的影响[J].眼科新进展,2014,34(4):326.[doi:10.13389/j.cnki.rao.2014.0088]
[12]朱恺,顾永昊,孙思勤. RNA干扰MMP-2基因对人晶状体上皮细胞增殖及移行能力的影响[J].眼科新进展,2015,35(3):235.[doi:10.13389/j.cnki.rao.2015.0063]
　ZHU Kai,GU Yong-Hao,SUN Si-Qin. Effects of RNA interference targeting MMP-2 0n proliferation and migration of human lens epithelial cells[J].Recent Advances in Ophthalmology,2015,35(5):235.[doi:10.13389/j.cnki.rao.2015.0063]
[13]杨静. 人原代晶状体上皮细胞中PEDF基因下调引起波形蛋白和αB-晶状体蛋白含量的变化[J].眼科新进展,2015,35(8):713.[doi:10.13389/j.cnki.rao.2015.0195]
　YANG Jing. Reduction of PEDF regulating expression of vimentin and αB-crystallin in human lens epithelial cells[J].Recent Advances in Ophthalmology,2015,35(5):713.[doi:10.13389/j.cnki.rao.2015.0195]
[14]曾新生,姚微微,彭燕一,等. d-δ-三烯生育酚对人晶状体上皮SRA细胞增殖和凋亡的影响[J].眼科新进展,2015,35(8):725.[doi:10.13389/j.cnki.rao.2015.0198]
　ZENG Xin-Sheng,YAO Wei-Wei,PENG Yan-Yi,et al. Effects of d-δ-tocotrienols on proliferation and apoptosis of human lens epithelial SRA cell line[J].Recent Advances in Ophthalmology,2015,35(5):725.[doi:10.13389/j.cnki.rao.2015.0198]

备注/Memo

备注/Memo:
 国家自然科学基金资助（编号：８１３７０９９７）
更新日期/Last Update: 2015-04-27