Guo Xin, Liu Cong, Wang Guo-Bing, Xu Ming-Guo
Zhuhai Campus, Zunyi Medical University, Zhuhai, Guangdong 519041, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2020 Jul;22(7):796-803. doi: 10.7499/j.issn.1008-8830.2001069.
To study the biomarkers for human coronary artery endothelial cell (HCAEC) injury induced by Kawasaki disease (KD) using isobaric tags for relative and absolute quantitation (iTRAQ) proteomics.
HCAECs cultured with the serum of children with KD were used as the KD group, and those cultured with the serum of healthy children was used as the healthy control group. The iTRAQ technique was used to measure the expression of proteins in two groups. The data on proteins were analyzed by bioinformatics. Western blot was used for the validation of protein markers.
A total of 518 significantly differentially expressed proteins were identified (with an absolute value of difference fold of >1.2, P<0.05). The gene ontology analysis showed that the differentially expressed proteins were significantly enriched in biological processes (including cellular processes, metabolic processes, and biological regulation), cellular components (including cell parts, cells, and organelles), and molecular functions (including binding, catalytic activity, and molecular function regulators). The KEGG analysis showed that the proteins were significantly enriched in the signaling pathways of ribosomes, PI3K-Akt signaling pathway, and transcriptional dysregulation in cancer. The PPI network showed that the top 9 protein markers in relation density were PWP2, MCM4, MCM7, MCM5, MCM3, MCM2, SLD5, HDAC2, and MCM6, which were selected as the protein markers for coronary endothelial injury in KD. Western blot showed that the KD group had significantly lower expression levels of the protein markers HDAC2, PWP2, and MCM2 than the healthy control group (P<0.05).
The serum of children with KD significantly changes the protein expression pattern of HCAECs and affects the signaling pathways associated with the cardiovascular system, which provides a new basis for the pathophysiological mechanism and therapeutic targets of KD.
采用相对和绝对定量同位素标记（iTRAQ）蛋白质组学技术研究川崎病（KD）诱导的人冠状动脉内皮细胞（HCAEC）损伤的生物标志物。
将用KD患儿血清培养的HCAEC作为KD组，用健康儿童血清培养的HCAEC作为健康对照组。采用iTRAQ技术检测两组蛋白质的表达情况。对蛋白质数据进行生物信息学分析。采用蛋白质免疫印迹法验证蛋白质标志物。
共鉴定出518种差异表达显著的蛋白质（差异倍数绝对值>1.2，P<0.05）。基因本体分析表明，差异表达的蛋白质在生物过程（包括细胞过程、代谢过程和生物调节）、细胞成分（包括细胞部分、细胞和细胞器）和分子功能（包括结合、催化活性和分子功能调节因子）中显著富集。KEGG分析表明，这些蛋白质在核糖体信号通路、PI3K-Akt信号通路和癌症中的转录失调信号通路中显著富集。蛋白质-蛋白质相互作用网络显示，关联密度排名前9的蛋白质标志物为PWP2、MCM4、MCM7、MCM5、MCM3、MCM2、SLD5、HDAC2和MCM6，这些被选为KD冠状动脉内皮损伤的蛋白质标志物。蛋白质免疫印迹法显示，KD组蛋白质标志物HDAC2、PWP2和MCM2的表达水平明显低于健康对照组（P<0.05）。
KD患儿血清显著改变HCAEC的蛋白质表达模式，并影响与心血管系统相关的信号通路，这为KD的病理生理机制和治疗靶点提供了新的依据。
