iTRAQ-based quantitative proteomics analysis of immune thrombocytopenia patients before and after Qishunbaolier treatment.
iTRAQ-based quantitative proteomics analysis of immune thrombocytopenia patients before and after Qishunbaolier treatment.
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DOI：
10.1002/rcm.8993
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发表时间：
2021-02-15
期刊：
影响因子：
--
通讯作者：
Bai H
中科院分区：
文献类型：
--
作者：
Burenbatu;Wang Y;Wang S;Narisu;Wuritunashun;Gong C;Hashengaowa;Eerdunduleng;Sarula;Guihua;Bai H
免疫性血小板减少症（ITP）的治疗通常包括长期使用免疫抑制皮质类固醇和脾切除术。然而，这些治疗通常对患者有副作用。蒙药芪顺宝利尔（QSBLE）疗效高，减少复发机会，无明显副作用。本研究旨在确定QSBLE治疗ITP的潜在治疗靶点。为了揭示QSBLE治疗前后ITP患者（ITPs）之间蛋白表达的差异，采用等压标签进行相对和绝对定量（iTRAQ）比较蛋白质组学研究。在正离子电喷雾电离模式下，采用纳米喷雾液相色谱/串联质谱（nano‐LC/MS/MS）进行分析。通过京都基因与基因组百科全书（KEGG）和其他生物信息学工具揭示了与ITP相关的关键蛋白。进行实时聚合酶链反应（RT - PCR）分析以确认差异表达蛋白。与对照组相比，在ITPs中共鉴定出982种差异表达蛋白。与QSBLE前处理组相比，QSBLE后处理组鉴定出61个差异表达蛋白，其中48个蛋白显著上调，13个蛋白下调。29条通路显著富集。Q6N030等蛋白在蛋白通路网络中起关键作用。进一步筛选了可能在ITP治疗中起重要作用的20种蛋白。根据蛋白质组学数据，RT‐PCR和Western blot分析进一步证实，QSBLE处理后ITPs中的MIF、PGK1和IGHM表达上调。相信这些鉴定的蛋白和生物信息学分析结果将为ITP治疗和生物标志物提供潜在的QSBLE治疗靶点。
Treatment of immune thrombocytopenia (ITP) usually involves long‐term use of immunosuppressive corticosteroids and splenectomy. However, these treatments often have side effects in patients. The Mongolian medicine Qishunbaolier (QSBLE) has a high curative effect, reduces the chances of relapse, and has no obvious side effects. This study was designed to identify potential therapeutic targets of QSBLE for treating ITP. To reveal differences in protein expression between ITP patients (ITPs) before and after QSBLE treatment, comparative proteomics studies were performed using isobaric tags for relative and absolute quantification (iTRAQ). The analysis used nanospray liquid chromatography/tandem mass spectrometry (nano‐LC/MS/MS) in positive ion electrospray ionization mode. Key proteins relevant to ITP were revealed by the Kyoto Encyclopedia of Genes and Genomes (KEGG) and other bioinformatics tools. Real‐time polymerase chain reaction (RT‐PCR) analysis was carried out for confirmation of differentially expressed proteins. A total of 982 differentially expressed proteins were identified in ITPs compared with the controls. Compared with the pre‐QSBLE treatment group, 61 differentially expressed proteins were identified in the post‐QSBLE treatment group, with 48 proteins being significantly upregulated and 13 downregulated. Twenty‐nine pathways were significantly enriched. Q6N030 and other proteins were the key players in the protein‐pathway network. Twenty proteins that may play important roles in the treatment of ITP were further filtered. RT‐PCR and Western blot analyses further confirmed that MIF, PGK1 and IGHM were upregulated in ITPs after QSBLE treatment, in accordance with the proteomics data. It is believed that the identified proteins and the results of bioinformatics analysis will provide a potential therapeutic target site for QSBLE for ITP therapy and biomarkers.
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