Proteomic temporal profile of human brain endothelium after oxidative stress.
Proteomic temporal profile of human brain endothelium after oxidative stress.
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DOI：
10.1161/strokeaha.110.585703
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发表时间：
2011-01
期刊：
影响因子：
8.3
通讯作者：
Lo EH
中科院分区：
文献类型：
--
作者：
Ning M;Sarracino DA;Kho AT;Guo S;Lee SR;Krastins B;Buonanno FS;Vizcaíno JA;Orchard S;McMullin D;Wang X;Lo EH
由于脑内皮细胞存在于神经血管界面，因此它们可以通过向循环中释放生物标志物来充当脑功能障碍的细胞报告者。我们使用蛋白质组学技术从人脑内皮培养物中筛选条件培养基，该培养物受到一氧化氮诱导的氧化应激超过 24 小时。通过酶联免疫吸附测定分析人类中风患者的血浆样本。在健康的内皮细胞中，相互作用图谱显示了涉及分泌因子、膜受体和基质成分的串扰。在受到氧化挑战的内皮细胞中，相互作用的蛋白质网络未能出现。相反，炎症标志物增加，分泌因子随时间振荡，内皮损伤修复表现为与基质完整性相关因子的变化。升高的炎症标志物包括热休克蛋白、趋化因子配体-1、血清淀粉样蛋白-A1、膜联蛋白-A5 和血小板反应蛋白-1。神经营养因子（prosaposin、nucleobindin-1 和速激肽前体）在 12 小时达到峰值，然后在 24 小时迅速下降。基底膜成分（纤连蛋白、桥粒糖蛋白、profiling-1）减少。细胞骨架标记物（肌动蛋白、波形蛋白、巢蛋白和细丝蛋白 B）随着时间的推移而增加。根据这一初步分析，我们在人血浆中进一步探索了高级候选血小板反应蛋白-1。与具有相似临床危险因素的对照组相比，急性缺血性卒中患者在症状出现 8 小时内的血小板反应蛋白-1 水平显着升高（659±81 vs 1132±98 ng/mL；P<0.05；n=20）。简化细胞培养系统的筛选可能有助于发现临床神经血管损伤的新型生物标志物。需要进一步的合作努力来发现和验证更多感兴趣的候选者。
Because brain endothelial cells exist at the neurovascular interface, they may serve as cellular reporters of brain dysfunction by releasing biomarkers into the circulation. We used proteomic techniques to screen conditioned media from human brain endothelial cultures subjected to oxidative stress induced by nitric oxide over 24 hours. Plasma samples from human stroke patients were analyzed by enzyme-linked immunosorbent assay. In healthy endothelial cells, interaction mapping demonstrated cross-talk involving secreted factors, membrane receptors, and matrix components. In oxidatively challenged endothelial cells, networks of interacting proteins failed to emerge. Instead, inflammatory markers increased, secreted factors oscillated over time, and endothelial injury repair was manifested as changes in factors related to matrix integrity. Elevated inflammatory markers included heat shock protein, chemokine ligand-1, serum amyloid-A1, annexin-A5, and thrombospondin-1. Neurotrophic factors (prosaposin, nucleobindin-1, and tachykinin precursors) peaked at 12 hours, then rapidly decreased by 24 hours. Basement membrane components (fibronectin, desomoglein, profiling-1) were decreased. Cytoskeletal markers (actin, vimentin, nidogen, and filamin B) increased over time. From this initial analysis, the high-ranking candidate thrombospondin-1 was further explored in human plasma. Acute ischemic stroke patients had significantly higher thrombospondin-1 levels within 8 hours of symptom onset compared to controls with similar clinical risk factors (659±81 vs 1132±98 ng/mL; P<0.05; n=20). Screening of simplified cell culture systems may aid the discovery of novel biomarkers in clinical neurovascular injury. Further collaborative efforts are warranted to discover and validate more candidates of interest.