基于iTRAQ标记结合IPA生物分析平台筛选异常黏液质型阳痿病证大鼠血清蛋白质组学研究

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目的 应用iTRAQ标记技术结合IPA生物分析平台,筛选异常黏液质型阳痿病证特异性差异表达蛋白候选标志物,并探讨其分子机制.方法 建立异常黏液质证候模型,并筛选阳痿病证模型后,分为正常组(N)、证候组(B1)和病证组(A1)共3组(n=10),分离血清,进行iTRAQ标记结合LC-MS-MS蛋白质组学分析,应用IPA在线生物平台对异常黏液质型阳痿病证组大鼠血清差异表达蛋白进行蛋白质调控网络分析、生物功能分析、典型通路分析和生物标志物筛查分析.结果 通过蛋白质组学技术获得61种异常黏液质型阳痿病证血清差异蛋白,IPA分析结果为,候选差异表达蛋白质中6种与血管内皮功能相关;3种与平滑肌活动相关;10种与炎症相关.参与的主要生物学过程为损伤反应、炎症反应和防御反应等,涉及的主要通路为LXR/RXR通路和FXR/RXR通路等,并获得与勃起功能障碍疾病相关的标志物4个,分别是C反应蛋白、血管紧张素原、T-激肽原1以及β-2-糖蛋白1.结论 炎症和血管内皮功能改变可能在该病证发生发展中发挥着关键性作用,上述4种蛋白可能可以作为异常黏液质型阳痿病证大鼠与勃起功能障碍疾病相关的血清候选差异蛋白.“,”Objective To screen the candidate serum biomarkers for abnormal phlegmatic syndrome with impotence disease using iTRAQ and IPA bioanalysis platform, and explore its molecular mechanism.Methods Rats with abnormal phlegmatic syndrome model were established, and abnormal phlegmatic syndrome rats model with impotence were selected and divided into three groups: the control group(N), phlegmatic syndrome model group(B1) and phlegmatic syndrome model group with impotence (A1) (n=10). Serum proteomics of these rats among three groups were comparatively analyzed by iTRAQ labeling combined with LC-MS-MS. Differentially expressed serum proteins were further analyzed by IPA online bioinformatics software,including networks analysis, biofunction Analysis, canonical Pathways and IPA Biomarker~Filter analysis.Results Total of 61 differentially expressed serum proteins were identified by iTRAQ. IPA bioanalysis showed that 6 different proteins were associated with vascular endothelial function, 3 proteins of smooth muscle activity,10 proteins associated with inflammation. The main biological processes of differential proteins were injury, inflammation reaction and defense response. The pathways involved in LXR/RXR activation and FXR/RXR activation. Four candidate biomarkers associated with Erectile Dysfunction disease were found, including C-reactive protein, Angiotensinogen, T-kininogen-1 andβ-2-glycoprotein 1.Conclusion Inflammation and vascular endothelial function may play a key role in the development of this disease, four proteins can be used as candidate proteins related to abnormal phlegmatic with impotence disease.
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