iTRAQ法分析孕期慢性应激子鼠脑海马组织中蛋白质组学变化  

The Proteomic Changes of Maternal Chronic Stress during Pregnancy on the Hippocampus of Offspring Based on iTRAQ Technology

作　　者：付有娟 关素珍 赵枫 刘红娅 陈小惠 戚发秋 刘志宏 FU Youjuan;GUAN Suzhen;ZHAO Feng;LIU Hongya;CHEN Xiaohui;QI Faqiu;LIU Zhihong(School of Public Health and Management,Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Environmental Factors and Chronic Disease Control,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院,银川750004 [2]宁夏环境因素与慢性病控制重点实验室,银川750004

出　　处：《宁夏医科大学学报》2021年第1期1-7,16,共8页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金(2018AAC02007)。

摘　　要：目的分析孕期慢性应激子鼠脑海马组织中差异表达的蛋白质及富集的信号通路,从整体水平上探讨可能的作用机制。方法通过慢性不可预知温和应激(CUMS)建立母鼠孕期慢性应激模型,模型组母鼠每天给予刺激,连续21 d;对照组母鼠正常环境饲养。应激前1天、第1、7、14、21天分别进行内眦静脉取血,测定母鼠血浆皮质酮含量。子鼠分为对照子鼠组和模型子鼠组,采用同位素标记的iTRAQ技术对子鼠海马组织的差异蛋白进行分析和鉴定,GO功能注释和KEGG富集分析差异蛋白参与的信号通路,String数据库分析差异蛋白间的相互作用。结果模型组母鼠血浆皮质酮含量高于同期对照组(P均<0.05),说明模型组大鼠处于慢性应激状态。两子鼠组共筛选和鉴定出5065个蛋白,差异蛋白有26个(P<0.05),上调蛋白19个(FC>1.2),下调蛋白7个(FC<0.83)。GO功能注释表明差异蛋白主要参与23种生物过程、14种细胞组分和9种分子功能。KEGG富集分析共鉴定到57条通路,其中显著富集的信号通路有8条(P<0.05),主要包括神经活性配体-受体相互作用、cGMP-PKG信号通路、胰腺分泌等。差异蛋白功能互作网络分析发现:酪氨酸蛋白激酶受体(INSR)、丝氨酸蛋白酶1(PRSS1)、cGMP依赖性蛋白激酶(PKG)、胆囊收缩素(CCK)关系最为密切。结论孕期慢性应激对子鼠脑海马发育的影响以神经活性配体-受体相互作用、cGMP-PKG信号通路及胰腺分泌为主的多种信号通路共同作用,与INSR、PRSS1、PKG、CCK等蛋白的差异表达密切相关。Objective To analyze the differential expressed proteins and enriched signal pathways in the hippocampus of pregnant rats with chronic stress,and to explore the possible mechanisms at the overall level.Methods The chronic stress model was established by chronic unpredictable mild stress(CUMS). The rats in the model group were stimulated daily for 21 days without interruption. The rats in the control group were fed in normal environment. Blood was collected from the angular vein one day before stress and at the first,7 th,14 th and 21 th days after stress,and the plasma corticosterone concentration of the mother rats was measured. The offspring rats were divided into control offspring group and model offspring group.By using the combination of iTRAQ technique to analyze and identify the differential proteins in the hippocampus of offspring rats,the signal pathway of differential proteins were analyzed by GO function annotation and KEGG enrichment,and the network of differential protein interaction between proteins was analyzed by string database. Results The content of plasma corticosterone in model group was significantly higher than that in control group(P all<0.05),indicating that the model group was in chronic stress state. The results of mass spectrometry showed that 5065 proteins were screened and identified in the two groups. There were 26 differential proteins(P<0.05),19 up regulating proteins(FC>1. 2),and 7 down regulating proteins(FC <0. 83). The results of GO function annotation showed that the differential proteins were mainly involved in 23 kinds of biological processes,14 cell components and 9 molecular functions. The results of KEGG enrichment analysis showed that there were 57 pathways,8 of which were significantly enriched(P<0. 05),mainly including the neuroactive ligand receptor interaction,cGMP-PKG signaling pathway,pancreatic secretion and so on. Based on the analysis of the interaction network of different protein functions in string database,it was found that the relationship among tyrosine p

关 键 词：iTRAQ技术 孕期慢性应激 子鼠 脑海马 蛋白质组学 

分 类 号：R714.2[医药卫生—妇产科学]