Transcellular distribution heterogeneity of Annexin A5 represents a protective response to lupus-related thrombophilia: a pilot Proteomics-based study.
Transcellular distribution heterogeneity of Annexin A5 represents a protective response to lupus-related thrombophilia: a pilot Proteomics-based study.
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DOI：
10.1016/j.bbrc.2012.02.162
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发表时间：
2012-04
影响因子：
3.1
通讯作者：
Di Zhou;N. Luo;Qiao Wu;Y. You;Z. Zhai;Zhirong Mou;Yuzhang Wu;F. Hao
中科院分区：
文献类型：
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作者：
Di Zhou;N. Luo;Qiao Wu;Y. You;Z. Zhai;Zhirong Mou;Yuzhang Wu;F. Hao
狼疮相关血管事件正成为系统性红斑狼疮（SLE）长期预后改善的巨大障碍，现有研究结果缺乏系统性。蛋白质组学是一种战略方法，但其在这方面的应用很少，主要涉及蛋白质组获取或生物标志物筛选，而不是功能鉴定。为了提供进一步的见解，我们研究了SLE患者外周血单个核细胞（PBMCs）的蛋白质组学多样性以及所鉴定的膜联蛋白A5 （AnxA5）在发病机制中的可能作用。该研究涉及214名SLE女性和183名健康女性。双向电泳凝胶图像显示，患者和对照组分别有649±25个和676±19个蛋白点。从这些蛋白点中选择了30个差异表达蛋白，并对其中的16个蛋白进行了质谱鉴定。Western blotting从患者的PBMCs中证实了过表达的候选物AnxA5 (SLE：对照=1.607:1，P=0.0004)，但elisa显示，与健康供者相比，患者血清中AnxA5的水平降低（SLE与对照=26.8±3.0 vs 49.0±3.3ng/mL， P<0.0001）。在寻找临床参数与患者血清两种分子水平的相关性时，发现血栓的表现与AnxA5呈正相关（Mann-Whitney Z=−2.084，P=0.037），而与抗AnxA5无关。SLE患者血浆凝血检测结果显示，升高的AnxA5可缩短凝血酶原时间，激活部分凝血活酶时间，延长凝血酶时间（P<0.001）。我们的数据显示了SLE患者和健康人之间PBMCs的蛋白质组学差异。此外，异质的跨细胞分布、细胞内浓度的升高和血清水平的降低代表了对狼疮相关的血栓形成的保护性反应；在SLE血栓形成过程中，AnxA5主要参与常见凝血途径。
Lupus-related vascular events are becoming a formidable obstacle to the improvement of long-term prognosis of systemic lupus erythematosus (SLE) and the existent findings lack for systematization. Proteomics is a strategic approach but its applications in this regard are rare and primarily involve proteome acquisition or biomarker screening, rather than functional identification. To provide further insight, we investigated the proteomic diversity of peripheral blood mononuclear cells (PBMCs) in SLE and the possible role of the identified Annexin A5 (AnxA5) in pathogenesis. The study involved 214 SLE and 183 healthy women. The two-dimensional electrophoresis gel images showed 649±25 and 676±19 protein spots from the PBMCs of the patients and controls, respectively. From these protein spots, 30 differentially expressed proteins were chosen, and 16 of these proteins were identified by mass spectrometer. Western blotting confirmed the over-expressed candidate, AnxA5, from the PBMCs of the patients (SLE:control=1.607:1, P=0.0004), but ELISAs indicated decreased levels of sera AnxA5 in the patients compared to healthy donors (SLE vs. control=26.8±3.0 vs. 49.0±3.3ng/mL, P<0.0001). A positive correlation was demonstrated between the manifestation of thrombosis and AnxA5 (Mann–Whitney Z=−2.084, P=0.037), not anti-AnxA5, while searching for correlations between clinical parameters and the two molecular levels of patient sera. The coagulation assays using plasma from SLE patients revealed that elevated AnxA5 could shorten prothrombin time, activated partial thromboplastin time and prolonged thrombin time (P<0.001). Our data demonstrated the proteomic differences in the PBMCs between SLE patients and healthy persons. Moreover, the heterogeneous transcellular distribution, increased intracellular concentrations and decreased serum levels of AnxA5 represent a protective response to lupus-related thrombophilia; AnxA5 mostly participate in the common coagulation pathway in the thrombogenesis of SLE.