孕期慢性应激致大鼠子代学习记忆能力损伤的蛋白质组学研究  

Proteomic study on the damage of learning and memory ability of rat offspring caused by chronic stress during pregnancy

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,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,宁夏环境因素与慢性病控制重点实验室,银川750004

出　　处：《中华劳动卫生职业病杂志》2021年第7期481-486,共6页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金地区基金(81960591);宁夏医科大学2018年校级科研基金项目(XT2018001)。

摘　　要：目的研究大鼠孕期慢性应激致子代学习记忆能力损伤相关的差异蛋白及信号通路,探讨可能的作用机制。方法于2019年7至10月,选择80~90日龄SPF级未曾受孕的雌性SD大鼠16只,体重(200±20)g;90~100日龄SPF级雄性SD大鼠12只,体重(220±20)g。适应性饲养1周后随机分组,雌鼠分为对照组和模型组(各8只);雄鼠分为对照交配组(8只)和模型交配组(4只),正常饲养。建立慢性不可预知温和应激(CUMS)模型,连续刺激21 d。应激前1天,第1、7、14、21天对雌鼠进行内眦静脉取血,测定皮质酮含量。利用Morris水迷宫实验检测子鼠的空间学习记忆能力,通过苏木素-伊红和尼氏染色观察子鼠海马组织形态学变化,采用同位素标记相对和绝对定量(iTRAQ)技术对子鼠海马组织进行蛋白质组学相关分析。结果与对照组比较,模型组雌鼠血浆皮质酮含量明显增高(F=7.717,P<0.05),模型建立成功。与对照子鼠组比较,模型子鼠组逃避潜伏期延长、游泳平均速度降低、跨越平台次数减少、目标象限游程缩短(P<0.05)。模型子鼠组海马组织结构中细胞形态不规则,神经元数量少,尼氏体分布不均匀、体积小、数量少。子鼠组共筛选出5065个蛋白,差异蛋白共26个(P<0.05),其中上调和下调分别为19、7个。差异蛋白主要参与23种生物学过程、14种细胞组分和9种分子功能。京都基因和基因组百科全书(KEGG)富集分析显示,共富集到57条通路,其中8条信号通路明显富集(P<0.05),可能参与子代学习记忆能力损伤的信号通路有5条,包括神经活性配体-受体相互作用、cGMP-PKG信号通路、黏附连接、FoxO信号通路和Notch信号通路,主要有酪氨酸蛋白激酶受体、α2A肾上腺素能受体、cGMP依赖性蛋白激酶等差异蛋白可能参与损伤过程。结论孕期慢性应激可能导致子代学习记忆能力损伤,蛋白质组学方法所富集的信号通路和筛选的关键差异蛋白可能在损Objective To study the differential protein and signal pathway related to the impairment of learning and memory ability of offspring caused by chronic stress during pregnancy and explore the possible mechanism.Methods From July to October 2019,sixteen SPF free female SD rats aged 80-90 days,weighing(200±20)g.Twelve SPF grade male SD rats aged 90-100 days,weighing(220±20)g.After a week of adaptive feeding,the female rats were randomly divided into control group and model group(8 rats in each group),male rats were divided into control mating group(n=8)and model mating group(n=4).Chronic unpredictable mild stress(CUMS)model was established and stimulated continuously for 21 days.One day before stress,the first,seventh,fourteenth and 21th day after stress,the blood was collected from the inner canthus vein of the female rats,and the content of corticosterone was determined.Morris water maze test was used to detect the spatial learning and memory ability of offspring rats.The morphological changes of hippocampus were observed by HE and Nissl staining.The proteomic correlation analysis of offspring rats'hippocampus was performed by isobaric tags for relative and absolute quantification(iTRAQ)technique.Results Compared with the control group,the content of plasma corticosterone in the model group was significantly higher(F=7.717,P<0.05),and the model was successfully established.In Morris water maze test,compared with the control offspring group,the escape latency was longer,the average swimming speed was lower,the number of crossing platform was less,and the target quadrant run was shorter in the model offspring group(P<0.05).The pathological results showed that the morphology of cells in the hippocampal tissue of the model offspring group was irregular,the number of neurons was small,Nissl body was unevenly distributed,the volume was small and the number was small.Mass spectrometry analysis showed that a total of 5065 proteins were screened out in the two offspring groups,and 26 proteins were differentially expresse

关 键 词：大鼠 孕期 慢性应激 学习记忆能力 子代 同位素标记相对和绝对定量技术 

分 类 号：R715.3[医药卫生—妇产科学]