Differential Proteomic Analysis of Syncytiotrophoblast Extracellular Vesicles from Early-Onset Severe Preeclampsia, using 8-Plex iTRAQ Labeling Coupled with 2D Nano LC-MS/MS
Differential Proteomic Analysis of Syncytiotrophoblast Extracellular Vesicles from Early-Onset Severe Preeclampsia, using 8-Plex iTRAQ Labeling Coupled with 2D Nano LC-MS/MS
复制标题
使用 8-Plex iTRAQ 标记结合 2D Nano LC-MS/MS 对早发严重先兆子痫的合体滋养层细胞外囊泡进行差异蛋白质组分析
DOI：
10.1159/000430283
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发表时间：
2015-01-01
影响因子：
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通讯作者：
Li, Li
中科院分区：
文献类型：
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作者：
Li, Hongmei;Han, Lei;Li, Li
目的：以往的研究表明，合体滋养层细胞外囊泡（STBM）脱落增加可能导致先兆子痫（PE）。我们的目的是确定STBM携带的蛋白质及其在早发性重度PE中的潜在病理作用。研究方法：在这项研究中，我们进行了差异蛋白质组学分析的STBM从早发性严重PE患者，使用iTRAQ同量异位素标签和二维纳米LC-MS/MS。STBM产生的体外外植体培养方法，然后通过电子显微镜和蛋白质印迹分析验证。结果：共鉴定出18 533个多肽和3 317个蛋白，定量分析了3 292个蛋白。我们在早发性重度PE患者STBM中鉴定了194个差异表达蛋白，其中122个蛋白表达上调，72个蛋白表达下调。进一步的生物信息学分析表明，跨膜转运子、跨膜转运蛋白和跨膜转运蛋白活性是基因本体（GO）注释中最丰富的类别。糖酵解/脂肪异生、柠檬酸循环、脂肪酸延伸、类固醇激素合成和氧化磷酸化是5条具有代表性的途径。采用蛋白质印迹法对4个与炎症、凝血或免疫调节相关的差异表达蛋白（siglec-6、calnexin、CD 63和S100-A8）进行了独立验证。结论：STBM携带的关键蛋白的鉴定不仅可以作为更好地理解和进一步探索PE的病因和发病机制的基础，而且还可以作为潜在的生物标志物，并为PE，特别是早发性重度PE（sPE）的未来治疗提供靶点。
Aims: Previous studies have revealed that the increased shedding of syncytiotrophoblast extracellular vesicles (STBM) may lead to preeclampsia (PE). We aimed to identify the proteins carried by STBM and their potential pathological roles in early-onset severe PE. Methods: In this study, we performed a differential proteomic analysis of STBM from early-onset severe PE patients, using iTRAQ isobaric tags and 2D nano LC-MS/MS. STBM were generated by the in vitro explant culture method, and then verified by electron microscopy and western blot analysis. Results: A total of 18 533 unique peptides and 3 317 proteins were identified, 3 292 proteins were quantified. We identified 194 differentially expressed proteins in STBM from early-onset severe PE patients, 122 proteins were up-regulated and 72 proteins were down-regulated. Further bioinformatics analysis revealed that mitochondrion, transmembrane transport and transmembrane transporter activity were the most abundant categories in gene ontology (GO) annotation. Glycolysis/ gluconeogenesis, citrate cycle, fatty acid elongation, steroid hormone biosynthesis and oxidative phosphorylation were the five significantly represented pathways. Four differentially expressed proteins (siglec-6, calnexin, CD63 and S100-A8) related to inflammation, coagulation or immunoregulation were independently verified using western blot. Conclusions: The identification of key proteins carried by STBM may serve not only as a basis for better understanding and further exploring the etiology and pathogenesis of PE, but also as potential biomarkers and in providing targets for future therapy in PE, especially in early-onset severe PE(sPE).