慢型克山病患者血清蛋白质双向凝胶电泳图谱分析

目的 分析克山病(KD)患者差异表达的血清蛋白质。 方法 以慢型KD和正常血清标本为研究对象,利用双向凝胶电泳(2-DE)分离KD和健康人血清蛋白质,银染显色,PowerLook2100XL扫描仪(Umax)透射扫描获取图像,ImageMaster 2D Platinum 5.0软件分析图像。并通过与ExPASy-SWISS-2DPAGE数据库关联,推测KD相关的差异蛋白。 结果 建立了稳定的慢型KD和健康人血清蛋白质2-DE图谱,分别检测到808和814个蛋白质点,匹配率96.5475%。软件分析显示2组样本共有44个差异蛋白质点,11个仅在KD组血清中表达,12个仅在正常血清组出现,21个为共有蛋白但在表达量上存在差异(KD组14个上调、7个下调,差异倍数 ≥ 3倍,P<0.01)。将差异蛋白与ExPASy-SWISS-2DPAGE数据库关联,在匹配值范围内出现的353个蛋白中,仅比对出KD 2-DE图谱胶内编号为1177的蛋白点在属性上与数据库中的P02774 2-D0004T6名为维生素D结合蛋白(VDBP)匹配度最高,推测编号为1177的差异蛋白是VDBP。 结论 KD患者与正常人血清蛋白质组存在明显差异。推测VDBP可能通过炎症免疫反应途径参与了KD的心肌损伤。

Two-dimensional Gel Electrophoresis Map of Serum Proteins in Patients with Chronic Keshan Disease

Objecitve To identify the different serum proteins expressed in patients with Keshan disease (KD). Methods Two -dimensional gel electrophoresis (2-DE) was performed with serum samples from the patients with chronic KD and healthy controls to separate serum proteins. The gels were stained by sliver and scanned by Umax scanner. The data were analyzed by ImageMaster 2D software. KD related proteins were identified through searching the ExPASy-SWISS-2DPAGE database. Results Stable two-dimensional gel electrophoresis maps were established for serum samples of KD patients and healthy controls. A total of 808 and 814 protein spots were observed in KD patients and healthy controls, respectively. The two maps had 96.5475% identical protein spots and 44 differentially expressed protein spots. Eleven protein spots were expressed exclusively in KD patients and 12 protein spots only appeared in healthy controls. About 21 proteins were expressed in both groups but varied in quantities (14 proteins were over-expressed by more than 3 times and 7 proteins were under-expressed by more than 3 times in KD patients, P<0.01). Among the 353 protein spots matched with the ExPASy-SWISS-2DPAGE databank, No.1177 protein appeared in the KD patient was found to have the closest match with P02774 2-D0004T6 known as vitamin D binding protein (VDBP). Conclusion There is a significant difference in serum protein expression between KD patients and normal people. VDBP might play a role in cardiac muscle damage via inflammatory immune reactions.