Endothelial cells microparticle-associated protein disulfide isomerase promotes platelet activation in metabolic syndrome.
Endothelial cells microparticle-associated protein disulfide isomerase promotes platelet activation in metabolic syndrome.
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内皮细胞微粒相关蛋白二硫键异构酶促进代谢综合征中的血小板活化
DOI：
10.18632/oncotarget.13081
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发表时间：
2016-12-13
期刊：
影响因子：
--
通讯作者：
Wang ZH
中科院分区：
文献类型：
--
作者：
Fan GQ;Qin RR;Li YH;Song DJ;Chen TS;Zhang W;Zhong M;Zhang Y;Xing YQ;Wang ZH
代谢综合征（MetS）是世界范围内常见的疾病，在MetS患者中血小板活化增强。然而，MetS中血小板活化的基本机制仍不完全清楚。内皮细胞损伤严重，释放出较多的内皮微粒（EMPs）。毕竟，emp是否参与血小板活化尚不清楚。如果是的话，它们是如何工作的？结果我们发现，在MetS患者中，EMPs、pmp（血小板衍生微粒）和微粒携带的pdi活性水平升高。IR内皮细胞释放更多emp， EMP-PDI更活跃。EMPs可以增强CD62P、GPIIb/IIIa和血小板聚集的激活，这一过程可以被RL90和芦丁等PDI抑制剂部分抑制。经EMPs刺激的活化血小板在细胞质上表达更多的PDI，释放更多的pmp。材料与方法我们采集了23例MetS患者和8例正常健康对照者的血浆。首先建立人脐静脉内皮细胞（HUVECs）胰岛素抵抗（IR）模型，然后从HUVECs培养基中分离EMPs，利用这些EMPs刺激血小板。流式细胞术检测微颗粒、p -选择素（CD62P）、糖蛋白IIb/IIIa （GPIIb/IIIa）水平，酶联免疫吸附试验（ELISA）检测EMPs水平。胰岛素转氢酶法检测蛋白二硫异构酶（PDI）活性。浊度法测定血小板聚集。结论EMPs可通过其表面携带的PDI促进血小板GPIIb/IIIa的活化和血小板聚集。
Background Metabolic syndrome (MetS) is a common challenge in the world, and the platelet activation is enhanced in MetS patients. However, the fundamental mechanism that underlies platelet activation in MetS remains incompletely understood. Endothelial cells are damaged seriously in MetS patients, then they release more endothelial microparticles (EMPs). After all, whether the EMPs participate in platelet activation is still obscure. If they were, how did they work? Results We demonstrated that the levels of EMPs, PMPs (platelet derived microparticles) and microparticle-carried-PDI activity increased in MetS patients. IR endothelial cells released more EMPs, the EMP-PDI was more activated. EMPs can enhance the activation of CD62P, GPIIb/IIIa and platelet aggregation and this process can be partly inhibited by PDI inhibitor such as RL90 and rutin. Activated platelets stimulated by EMPs expressed more PDI on cytoplasm and released more PMPs. Materials and Methods We obtained plasma from 23 MetS patients and 8 normal healthy controls. First we built insulin resistance (IR) model of human umbilical vein endothelial cells (HUVECs), and then we separated EMPs from HUVECs culture medium and used these EMPs to stimulate platelets. Levels of microparticles, P-selectin(CD62P), Glycoprotein IIb/IIIa (GPIIb/IIIa) were detected by flow cytometry and levels of EMPs were detected by enzyme-linked immunosorbent assay (ELISA). The protein disulfide isomerase (PDI) activity was detected by insulin transhydrogenase assay. Platelet aggregation was assessed by turbidimetry. Conclusion EMPs can promote the activation of GPIIb/IIIa in platelets and platelet aggregation by the PDI which is carried on the surface of EMPs.