Zhao Z L, Du S, Shen S X, Luo P, Ding S K, Wang G G, Wang L X
Department of Cardiology, Henan Provincial People's Hospital, Zhengzhou 450000, China.
Zhonghua Yi Xue Za Zhi. 2019 Jan 29;99(5):343-348. doi: 10.3760/cma.j.issn.0376-2491.2019.05.005.
To compare and analyze the differentially expressed plasma exosomic proteome between healthy control group (Control group) and viral myocarditis group (VMC group) to search for biomarkers that maybe used for early diagnosis of VMC. Fifty plasma samples of Control group and VMC group were collected respectively from Henan Provincial People's Hospital (from January 2016 to December 2017), and then 5 samples (1 ml) of each group were selected randomly, after exosomes extraction with ultra-centrifugation, difference gel electrophoresis (DIGE) was used to isolate the total proteins, and then the protein spots with more than 2-fold changes between VMC and Control group were picked up after the software analysis, afterward, the varied proteins were identified by MALDI-TOF/TOF mass spectrometry. Finally, the specifically related protein was selected to be verified by ELISA with the plasma exosomic samples of Control (40) and VMC (40). A total of 10 varied protein spots were found including 8 up-regulated proteins and 2 down-regulated proteins between VMC and Control group. After MS analysis, the up-regulated proteins in VMC group contained KRT2, KRT5, KRT9, KRT77, KRT78, AZGP1, HP and RBP4, whereas the down-regulated ones were CD5L and C1QB. RBP4 was selected to validate by ELISA analysis, and the corresponding results showed that RBP4 was increased specifically in plasma exosomes of VMC group (0.05) after comparing with Control group, which was consistent with DIGE. Ten proteins related to VMC are detected in total, and RBP4 might serve as a potential specific biomarker for early screening and diagnosis of VMC.
比较分析健康对照组(对照组)和病毒性心肌炎组(VMC组)之间血浆外泌体蛋白质组的差异表达,以寻找可能用于VMC早期诊断的生物标志物。分别从河南省人民医院收集50份对照组和VMC组的血浆样本(2016年1月至2017年12月),然后每组随机选取5份样本(1ml),经超速离心提取外泌体后,采用差异凝胶电泳(DIGE)分离总蛋白,软件分析后选取VMC组与对照组之间变化倍数超过2倍的蛋白点,随后通过基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF)鉴定差异蛋白。最后,选取特异性相关蛋白,用对照组(40例)和VMC组(40例)的血浆外泌体样本通过酶联免疫吸附测定(ELISA)进行验证。VMC组与对照组之间共发现10个差异蛋白点,包括8个上调蛋白和2个下调蛋白。质谱分析后,VMC组上调的蛋白包括角蛋白2(KRT2)、角蛋白5(KRT5)、角蛋白9(KRT9)、角蛋白77(KRT77)、角蛋白78(KRT78)、锌α2糖蛋白1(AZGP1)、触珠蛋白(HP)和视黄醇结合蛋白4(RBP4),而下调的蛋白为补体C1q亚成分B(C1QB)和补体d5配体(CD5L)。选取RBP4通过ELISA分析进行验证,相应结果显示,与对照组相比,VMC组血浆外泌体中RBP4特异性升高(P<0.05),与DIGE结果一致。共检测到10种与VMC相关的蛋白,RBP4可能作为VMC早期筛查和诊断的潜在特异性生物标志物。