合并与不合并2型糖尿病的下肢动脉硬化闭塞症患者血清差异性蛋白组学分析

Proteomics analysis on serum differential proteins between lower limb arteriosclerosis obliterans patients with and without type 2 diabetes mellitus

目的通过蛋白组学研究,鉴定合并2型糖尿病的下肢动脉硬化闭塞症(DLASO)患者与不合并2型糖尿病下肢动脉硬化闭塞症(LASO)患者血清的差异性蛋白,为防治2型糖尿病患者发生LASO提供依据。方法收集DLASO、LASO各15例患者的血清,对蛋白进行提取、定量和酶解,应用同位素标记相对和绝对定量(iTRAQ)结合二维液相色谱串联质谱(2DLC-MS/MS)技术,鉴定DLASO、LASO患者血清中的蛋白,筛选出DLASO患者与LASO患者的差异表达蛋白。利用PANTHER数据库对鉴定得到的差异性蛋白进行生物学过程、分子作用和细胞组分分析。结果共鉴定出蛋白质338种,筛选出差异性蛋白27种,其中DLASO患者出现表达上调蛋白8种、表达下调蛋白19种。在生物学过程方面,差异性蛋白主要为参与细胞反应(48.1%)、代谢过程(48.1%)和生物黏附(29.6%)的蛋白;在分子作用方面,差异性蛋白主要为参与细胞结合作用(33.3%)、催化活性作用(29.6%)和结构分子活性作用(18.5%)的蛋白;在细胞组分方面,差异性蛋白主要为位于细胞区域(14.8%)、细胞器(14.8%)和细胞外区域(14.8%)的蛋白。结论发现DLASO与LASO患者血清中的差异性蛋白27种,这些差异性蛋白可作为防治DLASO的潜在生物学靶点。

Objective To identify the differentially expressed proteins between lower limb arteriosclerosis obliterans patients with type 2 diabetes mellitus（ DLASO） and patients without type 2 diabetes mellitus（ LASO） by the proteomic research. Methods Serum samples were separately collected from DLASO patients and LASO patients（ 15 cased in each group）. Isobaric tags for relative and absolute quantitation（ iTRAQ）-tagging combined with two-dimensional liquid chromatography-tandem mass spectrometry（ 2D LC-MS/MS） analysis were used to identify the differentially expressed proteins between DLASO patients and LASO patients. The differentially expressed proteins were analyzed for biological processes,molecular effects and cell composition by using PANTHER database. Results Compared with LASO patients,a total of 27 differentially expressed proteins were identified in DLASO patients,8 proteins were significantly up-regulated while 19 proteins were significantly down-regulated. Biological process analysis showed that the differential protein was mainly involved in cell reaction（ 48. 1%）,metabolic process（ 48. 1%）,and bioadhesion（ 29. 6%）. The results of molecular analysis showed that the differential protein was mainly involved in cell binding（ 33. 3%）,catalytic activity（ 29. 6%）,and structural molecule activity（ 18. 5%）. Cell composition analysis showed that the differential proteins were mainly located in the cell region（ 14. 8%）,organelles,（ 14. 8%） and extracellular regions（ 14. 8%）. Conclusion Totally 27 differentially expressed proteins between DLASO and LASO patients are identified by proteomic research,which may be used as potential biological targets for the prevention and treatment of DLASO.