微波辐射后晶状体上皮细胞与未辐射晶状体上皮细胞蛋白质组双向电泳图谱比较

Two-dimensional electrophoregram for proteomic analysis of human lens epithelial cells under microwave and control

关键词:  人晶状体上皮细胞  微波辐射  双向电泳  蛋白质组

Key words:  human lens epithelial cells  microwave radiation  two dimensional electrophoresis  proteome

摘要:

［目的］ 比较正常晶状体上皮细胞与微波辐射后晶状体上皮细胞蛋白质组双向电泳图谱差异，从蛋白质组水平初步探索微波辐射对人眼晶状体的损伤。［方法］体外培养人晶状体上皮细胞株，随机分为实验组和对照组，实验组用SAR值为4.0 W/kg的1800 MHz制式微波辐照2 h，对照组同一辐射箱培养但不辐射。辐照后立即提取总蛋白质，固相pH梯度(IPG)等电聚焦双向凝胶电泳进行蛋白质分离，银染显色的凝胶通过GS-800扫描仪获取图像，使用PDQuest专业图像分析软件分析电泳图像。［结果］正常晶状体上皮细胞与微波辐射后晶状体上皮细胞分别检出897个和981个蛋白质斑点。对2张电泳图进行匹配后，发现有4个蛋白质点在辐射后细胞蛋白质组图谱中上调表达，3个蛋白质点下调表达。［结论］初步建立了人晶状体上皮细胞比较蛋白质组学的实验方法；差异蛋白质的发现为深入理解辐射性白内障发病机制提供了有益的线索。

Abstract:

［Objective］To investigate the molecular mechanisms of radiation cataract by comparing global protein patterns in two-dimensional electrophoregram of the human lens epithelial cells (HLECs) exposed and sham-exposed to microwave.［Methods］Cultured human HLECs were randomly divided into experimental and control groups. They were exposed and sham-exposed to 1800 MHz GSM-like radiation at 4.0 W/kg for 2 hours in an sXc-1800 incubator and irradiate system to give well controlled atmospheric conditions. On the moment of the radiation accomplished, proteome was extracted and then separated using immobilized pH gradients(IPG) two-dimensional electrophoresis and visualized by silver staining. The digitized images which got with GS-800 scaner（Bio-Rad） were then analyzed with PDQuest software in order to establish the differential expression profiles.［Results］On average, 981 and 897 protein spots could be obtained in the electrophoregraphs for HLECs under microwave and the control. Four of this spots were detected as up-regulated and 3 were down-regulated after radiation. ［Conclusions］The differentially displayed proteins in the HLECs identified with microwave and control may provide further insight into the radiation cataract.