妊娠期绵羊缺氧对胎羊心肌组织低氧反应蛋白表达及信号通路的影响  

Effects of hypoxia in pregnant sheep on expression and signaling pathway of hypoxia-responsive proteins in fetal sheep myocardium

作　　者：胡劼 焦明克 李锐 李华[3] Hu Jie;Jiao Mingke;Li Rui;Li Hua(Department of Ultrasound Medicine,Yubei Hospital,Chongqing 401120,China;Department of Ultrasound Medicine,The Third Affiliated Hospital of Chongqing Medical University,Chongqing 401120,China;Department of Cardiac Ultrasound,Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,China)

机构地区：[1]重庆市渝北区人民医院超声医学科,重庆401120 [2]重庆医科大学附属第三医院超声医学科,重庆401120 [3]新疆医科大学附属中医医院心脏超声科,乌鲁木齐830000

出　　处：《中华航海医学与高气压医学杂志》2025年第2期158-163,共6页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：国家自然科学基金(81560291)。

摘　　要：目的:探讨妊娠期绵羊缺氧对胎羊心肌组织低氧反应蛋白表达及信号通路的影响。方法:按照随机数字表法将30只健康妊娠期绵羊分为低海拔对照(LAC)组、轻度缺氧暴露(MHE)组和重度缺氧暴露(SHE)组,每组10只。MHE组、SHE组分别于模拟海拔3 000、5 000 m环境的人工实验舱内饲养,LAC组于平原环境实验舱内饲养,持续整个妊娠期直至分娩前。提取胎羊心肌组织蛋白后,应用高效液相色谱系统分离蛋白,采用Q-Exactive质谱仪进行质谱分析,利用内嵌Proteome Discoverer 1.4的Mascot 2.2软件进行蛋白种类鉴定,根据离子峰强度值对肽段进行定量分析。通过Heat mapper server对蛋白进行分层聚类,应用geneontology进行基因分析,根据所鉴定蛋白的基因注释和生物学功能对其分子功能进行分类。利用KEGG通路数据库分析蛋白的代谢途径,通过STRING程序分析蛋白相互作用的网络。结果:2个缺氧暴露组与LAC组比较,共存在284种差异蛋白(160种蛋白表达上调,124种蛋白表达下调);MHE组与SHE组比较,共存在197种差异蛋白(80种蛋白表达上调、117种蛋白表达下调)。3组心肌组织共有80种低氧反应蛋白,其中41种蛋白表达上调、39种蛋白表达下调。聚类树结果显示,低氧条件下所有低氧反应蛋白的表达水平明显下降,缺氧对蛋白代谢有抑制作用。低氧反应蛋白涉及最多的生物过程是翻译过程,明显富集的细胞成分为溶酶体膜、细胞膜和高尔基体,分子功能为染色质结合、相同蛋白结合和锌离子结合,主要影响甘油磷脂代谢、蛋白消化吸收和松弛素信号通路。与LAC组相比,2个缺氧暴露组心肌组织中的低氧反应蛋白存在明显表达调节,并且存在相互作用的蛋白网络。结论:妊娠期绵羊缺氧对胎羊胚胎心脏发育过程中的低氧反应蛋白表达及信号通路均产生明显影响。Objective:To explore the effects of hypoxia in pregnant sheep on the expression and signaling pathway of hypoxia-responsive proteins in fetal sheep myocardium.Methods:A total of thirty healthy pregnant sheep were divided into low-altitude control(LAC)group,mild hypoxia exposure(MHE)group,and severe hypoxia exposure(SHE)group,according to the random number table method,with 10 sheep in each group.The MHE and SHE groups were housed in the artificial experimental chambers simulating altitude of 3000 m and 5000 m,respectively;while the LAC group was kept in a plain environment chamber throughout the whole pregnancy until just before delivery.Myocardial proteins from fetal sheep myocardial tissue were extracted,and then isolated by a high-performance liquid chromatography system.After that,Q-Exactive mass spectrometer was used for mass spectrometry analysis,Mascot 2.2 software embedded in Proteome Discoverer 1.4 was used to identify the protein species,and the peptide segment was quantitatively analyzed according to the ion peak intensity value.Proteins were stratified and clustered using Heat mapper server,and gene analysis was carried out by gene ontology.Molecular functions of identified proteins were classified based on gene annotation and biological functions.Protein metabolic pathway was analyzed using KEGG pathway database,and protein-protein interaction network was examined using STRING's website.Results:Compared with LAC group,there were 284 kinds of differential proteins(160 up-regulated and 124 down-regulated)in the two hypoxic exposure groups.There were 197 kinds of differential proteins between the MHE group and the SHE group(80 up-regulated and 117 down-regulated).There were 80 kinds of hypoxia-responsive proteins in the myocardium of the 3 groups,with 41 up-regulated and 39 down-regulated.The cluster tree results showed a significant decrease in the expressions of all hypoxia-responsive proteins under hypoxic conditions,indicating an inhibitory effect of hypoxia on protein metabolism.The most involved b

关 键 词：妊娠期 缺氧 心血管发育 蛋白质谱分析 绵羊 

分 类 号：R73[医药卫生—肿瘤]