肺组织灌注前后双向电泳的比较  

Comparison of 2-Dimensional electrophoresis of pulmonary tissue proteins before and after saline perfusion

作　　者：李圣青[1] 赵峰[1] 戚好文[1] 张晓君[2] 赵馨[2] 潘刚[2] 吴哲[3] 王宇[3] 阙海萍[2] 刘少君[2] 

机构地区：[1]第四军医大学西京医院呼吸科,陕西西安710033 [2]解放军军事医学科学院三所一室,北京100850 [3]吉林大学生命科学院,吉林长春130021

出　　处：《西安交通大学学报（医学版）》2006年第2期203-206,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的寻找更加有效的双向电泳方法以展示肺组织蛋白。方法取6只Wistar大鼠随机分为对照组和灌注组,用丙酮/三氯醋酸法提取肺组织蛋白并进行双向电泳分离,将2-D胶银染后用ImageScanner扫描仪扫描,采用ImageMaster 2-D Elite 3.10软件进行图象分析。结果对照组2-D胶的蛋白点计数为1 095±5,灌注组蛋白点计数为1 569±10。在分子质量30-70 ku范围内,灌注组的蛋白点计数显著高于对照组(P<0.01),在其他分子质量范围内二者蛋白点计数无显著性差异。在pH值5-8、9-10范围内,灌注组的蛋白点计数显著高于对照组(P<0.01),在其他pH值范围内二者蛋白点计数无显著性差异。结论生理盐水灌注法可显著减少肺组织中的血液蛋白成分,使得肺组织蛋白可以更好的在2-D胶上展示。Objective To find a better way to display lung tissue proteins by 2-dimensional electrophoresis （2-DE）. Methods Lung tissue was obtained directly or after saline perfusion. Proteins were extracted by acetone/ TCA methods and separated by 2-DE to identify differently displayed proteins. The silverstained 2-DE were scanned with digital ImageScanner and analyzed with ImageMaster 2-D Elite 3.10 software. Comparison of protein spots obtained directly or after saline perfusion was carried out in statistical ways. Results The saline perfusion group displayed 1 569±10 protein spots on 2-DE gels and the control group 1 095±5 protein spots. The saline perfusion group displayed more proteins between 30-70 ku than the control group, and had no difference with it in the other molecular weight range. The saline perfusion group also displayed more proteins between pH 5-8 and 9-10 than the control group, and had no difference with it in the other pH range. Conclusion The saline perfusion methods can greatly reduce a few families of high abundance proteins（albumin, immunoglobulins） in sera, so the lung tissue proteins can be better displayed on 2-DE gels.

关 键 词：双向电泳 肺组织 灌注 

分 类 号：R563.5[医药卫生—呼吸系统]