中医不同证型乳腺增生患者血清蛋白质组的研究

目的：应用凝胶内差异显示电泳和质谱技术研究不同证型乳腺增生病患者血清蛋白质组。方法：收集正常人及不同证型乳腺增生病患者血清,去除高丰度蛋白质后分别用Cy3或Cy5标记,每一对Cy3和Cy5标记样品都与一个Cy2标记的内标等量混合,上样于同一胶中进行电泳分离,经不同光激发下扫描得到不同样品的蛋白质组图谱。所获得的图谱经DeCyder6.5软件进行分析,筛选表达量有显著差异的蛋白质进行质谱鉴定。结果：与正常人相比,在乳腺增生患者血清中抗凝血酶lii、富含亮氨酸的α-2糖蛋白、HCCR结合蛋白2、结合珠蛋白2和转甲状腺蛋白及其变异体表达量增加。而SP40,40、Ras association and pleckstrin homology domains 1 iso-form3表达量下降。在不同证型乳腺增生患者血清中,抗凝血酶lii和SP40,40在肝郁气滞型表达量最高,痰瘀互结型次之,而冲任失调型表达量少;HCCRBP2在痰瘀互结型中表达量最高,在肝郁气滞型中表达量最低;转甲状腺素蛋白变异体在肝郁气滞型、冲任失调型和痰瘀互结型中表达依次降低。结论：这些蛋白质可能与乳腺增生病不同证型相关,可以作为乳腺增生病临床中医辨证论治的候选客观指标。

Serum Proteome Study on Hyperplasia of Mammary Glands with Different Syndrome

Objective：Two-dimensional difference gel electrophoresis and mass spectrum were used to study the serum proteome of hyperplasia of mammary glands with different syndrome.Methods：The serum of normal people and patients with different syndromes hyperplasia of mammary glands were collected,and labeled with Cy3 or Cy5 randomly after removed high abundance proteins. Each Cy3-labeled sample and Cy5-labeled sample was mixed on the same 2-D gel along with a Cy2-labeled mixture of all samples as an internal standard. The gels were respectively imaged by Typhoon 9400 using different emission filter and spot-features were analyzed by DeCyder 6.5 software,and the different proteins were identified with mass spectrum.Results：Compared with normal group,in the serum of patients with breast hyperplasia antithrombin lii,leucine-rich α-2 glycoprotein,HCCRBP2,haptoglobin and transthyretin and its variants express increased. The SP40,40,Ras association and pleckstrin homology domains 1 isoform 3 expression decreased. In different syndromes,the expression of anti-thrombin lii and SP40,40 in Ganyu Qizhi syndrome is highest,followed by Tanyu Hujei syndrome,while the Chongren Shitiao syndrome expression less; HCCRBP2 expressed highest in Tanyu Hujei syndrome,and the lowest in Ganyu Qizhi syndrome; The expression of transthyretin variants in Ganyu Qizhi syndrome,Chongren Shitiao syndrome,Ganyu Qizhi syndrome in turn reduced. Conclusion：These proteins might be related to hyperplasia of mammary glands with different syndrome,and could become candidate marks for clinic differentiation of syndromes in hyperplasia of mammary glands.