Microarray-based analysis of gene expression profiles in peripheral blood of patients with acute primary angle closure

Background: We investigated the expression of molecules in peripheral blood mononuclear cells (PBMCs) and plasma of patients with acute primary angle closure (APAC). Materials and methods: Peripheral blood was collected from patients with APAC (n = 10) and age-matched controls (n = 5). The gene transcription profile was analyzed in PBMCs using microarrays and validated by real-time reverse transcription polymerase chain reaction (RT-PCR). The levels of secreted proteins were evaluated in plasma by ELISA. Results: 347 gene transcripts were up-regulated by 2-fold or more, and 696 transcripts down-regulated 2-fold or more in PBMCs from patients compared to controls. The most highly up-regulated gene was thrombospondin-1 (TSP-1, 8.66-fold increase), and the most down-regulated gene was prostaglandin-endoperoxide synthase 2 (PTGS2, 9.09-fold decrease). Real-time RT-PCR assay confirmed the increase of TSP-1 and the decrease of PTGS2 in PBMCs of patients. ELISA revealed that the levels of TSP-1 and active transforming growth factor (TGF)-β1 that is activated by TSP-1 were elevated in plasma of patients, while the level of prostaglandin E2 (PGE2) that is converted by PTGS2 was reduced. The plasma level of TSP-1 was positively correlated with that of active TGF-β1. Conclusions: Our data suggest that the molecular network including TSP-1, TGF-β1, and PGE2 might be involved in the pathogenesis of APAC and PACG.

研究背景：本研究旨在探讨急性原发性闭角型青光眼（acute primary angle closure, APAC）患者外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）及血浆中的分子表达特征。材料与方法：纳入10例APAC患者与5例年龄匹配的健康对照者，采集外周血样本。采用基因芯片技术分析PBMCs的基因转录谱，并通过实时逆转录聚合酶链反应（real-time reverse transcription polymerase chain reaction, RT-PCR）进行验证；采用酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）检测血浆中分泌蛋白的水平。研究结果：与对照组相比，APAC患者PBMCs中共有347个基因转录本上调2倍及以上，696个基因转录本下调2倍及以上。其中上调幅度最高的基因为血小板反应蛋白-1（thrombospondin-1, TSP-1，上调8.66倍），下调幅度最高的基因为前列腺素内过氧化物合酶2（prostaglandin-endoperoxide synthase 2, PTGS2，下调9.09倍）。实时RT-PCR验证结果证实，患者PBMCs中TSP-1表达上调、PTGS2表达下调。ELISA检测显示，患者血浆中TSP-1及TSP-1激活的活性转化生长因子β1（transforming growth factor-β1, TGF-β1）水平升高，而PTGS2催化生成的前列腺素E2（prostaglandin E2, PGE2）水平降低；且患者血浆TSP-1水平与活性TGF-β1水平呈正相关。研究结论：本研究数据提示，包含TSP-1、TGF-β1及PGE2的分子网络可能参与APAC及原发性闭角型青光眼（PACG）的发病机制。