肺组织灌注前后双向电泳的比较

　　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2D胶银染后用ImageScanner扫描仪扫描，采用ImageMaster 2D Elite 3.10软件进行图象分析。结果 对照组2D胶的蛋白点计数为1095±5，灌注组蛋白点计数为1 569±10。在分子质量30-70ku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2D 胶上展示。

　　关键词：双向电泳;肺组织;灌注

　　Comparison of 2Dimensional electrophoresis of pulmonary　tissue proteins before and after saline perfusionLi Shengqing, Zhao Feng, Qi Haowen, Zhang Xiaojun, Zhao Xin,Pan Gang, Wu Zhe, Wang Yu, Que Haiping, Liu Shaojun

　　(1. Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xian 710032;

　　2. Institute of Basic Medical Sciences, Academy of Military Medical Sciences,Beijing 100850;

　　3. Academy of life Sciences, Jilin University, Changchun 130021, China)

　　ABSTRACT: Objective To find a better way to display lung tissue proteins by 2dimensional electrophoresis(2DE). Methods Lung tissue was obtained directly or after saline perfusion. Proteins were extracted by acetone/TCA methods and separated by 2DE to identify differently displayed proteins. The silverstained 2DE were scanned with digital ImageScanner and analyzed with ImageMaster 2D 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 1569±10 protein spots on 2DE gels and the control group 1095±5 protein spots. The saline perfusion group displayed more proteins between 30-70ku 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 protei

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