特发性高草酸尿大鼠肝脏蛋白质组的电泳分析

Proteome Study of the Liver of Rats with Idiopathic Hyperoxaluria by Two-Dimensional Difference Gel Electrophoresis

目的:建立稳定的特发性高草酸尿大鼠肝脏蛋白质组的双向电泳图谱,探讨其差异蛋白质组与特发性高草酸尿的关系。方法:取特发性高草酸尿大鼠及正常对照大鼠的肝组织300 mg,匀浆提取肝组织总蛋白,分别用Cy3或Cy5标记,每一对Cy3和Cy5标记样品均与一个Cy2标记的内标等量混合,采用凝胶内差异显示电泳(DIGE)技术进行电泳分离,经过不同激光下扫描得到不同样品的蛋白质组图谱。获得的图谱经DeCyderTMv6.5软件进行分析。结果:在特发性高草酸尿大鼠肝组织中共筛选出21个差异表达蛋白质点,有11个蛋白质表达水平显著增加,另外10个蛋白质表达水平显著下降。结论:利用DIGE技术可以作胶内对比分析,并可根据内标消除胶与胶之间的差异,从而提高统计可信度。分析出的21个差异蛋白质可能与特发性高草酸尿的发生有密切关系。

Objective： To establish a stable two-dimensional electrophoresis map of idiopathic urinary oxalate liver proteome, to explore the relationship between the differential proteomics and the idiopathic urinary oxalate. Methods：The liver of rats with idiopathic hyperoxaluria and the normal liver 300 mg tissue were homogenized with lysis solution. The extracts were labeled with Cy3 or CyS. Each Cy3-1abeled sample and CyS-labeled sample were mixed on the same 2-D gel along with a Cy2-1abeled mixture of all samples as an internal standard. The gels imaged by Typhoon scanner using different emission filter and spot were analyzed by DeCyderTM v6.5 software in each comparison. IR^lts：Intensity changes of 21 spots were detected with statistical significance. 11 proteins of them were changing in abundance more than 1.5 folder up while 10 proteins decreased more than 1.5 folder down. C.onehmio~：Using DIGE technology with internal standard, statistically significant quantitative comparisons of each protein abundance change could be made across multiple samples simultaneously without interference due to gel to gel variation. And these 21 proteins might be involved in the process of idiopathic hyperoxaluria.