肺组织灌注前后双向电泳的比较

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。

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

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.