基于iTRAQ技术对化疗所致认知障碍机制的研究

Application of iTRAQ Technology in the Exploration of the Mechanism of Nab-Paclitaxel-Induced Cognitive Impairment

目的基于放射性核素标记相对与绝对定量技术(isobaric tags for relative and absolute quantitation,iTRAQ)探究化疗相关认知障碍(chemotherapy-induced cognitive impairment,CICI)的发生机制。方法雄性SD大鼠6只,随机分为对照组和化疗组,化疗组大鼠静脉注射环磷酰胺和阿霉素,每周1次,共3周;对照组大鼠给予等剂量的0.9%NaCl注射液。通过iTRAQ技术探究两组间的差异蛋白,根据GO和KEGG数据库分析对差异蛋白的生物功能及相关信号通路进行富集。结果蛋白质组学共鉴定出52764条肽段和7670个蛋白,化疗组与对照组的差异蛋白为434种,其中显著下调蛋白338种,显著上调蛋白96种。结果显示差异蛋白主要富集于细胞器、细胞膜等细胞成分;主要参与细胞代谢、生物调节及代谢等生物学过程;显著富集于结合、催化等分子功能。对差异蛋白进行KEGG信号通路富集分析显示其主要富集于单纯疱疹病毒感染、免疫排斥、自身免疫性甲状腺疾病等与免疫相关的通路。通过构建蛋白-蛋白相互作用(protein-protein interaction,PPI)网络发现信号转导及转录激活因子(signal transducer and activator of transcription,STAT)及泛素样修饰剂ISG15处于中心位置,均与免疫相关。结论本研究利用iTRAQ技术CICI的可能机制进行探究,结果进一步说明CICI可能与神经炎症有关,特别是其与自身免疫的相关性值得进一步研究。γ干扰素(interferonγ,IFN-γ)可能是CICI潜在的生物学标志物及治疗靶点。

Objective To investigate the mechanism of chemotherapy-induced cognitive impairment(CICI)based on the isobaric tags for relative and absolute quantitation(iTRAQ)technique.Methods Six male SD rats were randomly divided into chemotherapy group and control group.Rats in the chemotherapy group were injected intravenously cyclophosphamide(40mg/kg)and doxorubicin(4mg/kg)once a week for 3 weeks,and the control group was given the same amount of saline.The differential protein profiles between the two groups were explored by the iTRAQ,and the biological functions of the differential proteins and the signaling pathways involved were analyzed according to the GO and KEGG databases.Results A total of 52764 peptides and 7670 proteins were identified by proteomics,and 434differential proteins(338significantly down-regulated proteins and 96significantly up-regulated proteins)were identified between the chemotherapy group and the control.The results showed that the differential proteins were mainly enriched in cellular components such as organelles and cell membranes;biological processes mainly included cellular metabolic processes,bioregulation,and metabolic processes;molecular functions such as binding and catalysis were significantly enriched.The KEGG signaling pathway enrichment analysis showed that the differential proteins were mainly enriched in immune-related pathways,such as herpes simplex infection,autoimmune thyroid disease,and allograft rejection.The construction of PPI revealed that STAT and ubiquitin-like modifier ISG15 were centrally located and both were associated with immunity.Conclusion In this study,the possible mechanisms of CICI were investigated by iTRAQ technology further suggests that CICI may be related to neuroinflammation,especially in autoimmunity.IFN-γmay be a potential biomarker and therapeutic target for CICI.