Wang Yanbo, Wang Shuanglian, Gong Cuiqin, Bai Haihua
Affiliated Hospital of Inner Mongolia University for Nationalities, Tongliao, Inner Mongolia, 028000, China.
College of Life Sciences and Food Sciences, Inner Mongolia University for Nationalities, Tongliao, Inner Mongolia, 028000, China.
Rapid Commun Mass Spectrom. 2021 Feb 15;35(3):e8993. doi: 10.1002/rcm.8993.
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.
免疫性血小板减少症（ITP）的治疗通常涉及长期使用免疫抑制性皮质类固醇和脾切除术。然而，这些治疗方法在患者中常常有副作用。蒙药七顺宝力尔（QSBLE）具有较高的疗效，降低复发几率，且无明显副作用。本研究旨在确定QSBLE治疗ITP的潜在治疗靶点。
为揭示QSBLE治疗前后ITP患者（ITPs）蛋白质表达的差异，使用相对和绝对定量的等压标签（iTRAQ）进行了比较蛋白质组学研究。分析采用正离子电喷雾电离模式的纳喷雾液相色谱/串联质谱（nano-LC/MS/MS）。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与ITP相关的关键蛋白质。进行实时聚合酶链反应（RT-PCR）分析以确认差异表达的蛋白质。
与对照组相比，ITPs中共鉴定出982种差异表达蛋白质。与QSBLE治疗前组相比，QSBLE治疗后组鉴定出61种差异表达蛋白质，其中48种蛋白质显著上调，13种下调。29条通路显著富集。Q6N030等蛋白质是蛋白质-通路网络中的关键参与者。进一步筛选出20种可能在ITP治疗中起重要作用的蛋白质。RT-PCR和蛋白质免疫印迹分析进一步证实，QSBLE治疗后ITPs中MIF、PGK1和IGHM上调，与蛋白质组学数据一致。
据信，鉴定出的蛋白质和生物信息学分析结果将为QSBLE治疗ITP提供潜在的治疗靶点位点和生物标志物。
