基于4D蛋白组学分析RhoE基因敲除对糖尿病大鼠心脏组织中蛋白质表达的影响及机制  

Effect and mechanism of knockout RhoE on the protein expression profiles in cardiac tissues of diabetic rats based on 4D proteomics

作　　者：赵鋆 施凯佳 伍彩霞 邹园 吴林栩 陈旭 申志华 郭峻莉[1] 揭伟 ZHAO Yun;SHI Kai-jia;WU Cai-xia;ZOU Yuan;WU Lin-xu;CHEN Xu;SHEN Zhi-hua;GUO Jun-li;JIE Wei(Hainan Provincial Key Laboratory of Tropical Cardiovascular Disease Research,the First Affiliate Hospital,Hainan Medical University,Haikou 571199,China;Department of Pathology,School of Basic Medicine,Guangdong Medical University,Zhanjiang 524023,China;Department of Endocrinology,the First Affiliated Hospital,Hainan Medical University,Haikou 571199,China)

机构地区：[1]海南医学院第一附属医院海南省热带心血管病研究重点实验室,海南海口571199 [2]广东医科大学基础医学院病理学系,广东湛江524023 [3]海南医学院第一附属医院内分泌科,海南海口571199

出　　处：《海南医学院学报》2022年第23期1761-1770,1778,共11页Journal of Hainan Medical University

基　　金：国家自然科学基金项目(82060053);海南省重点研发社会发展专项(ZDYF2020122)。

摘　　要：目的:分析RhoE基因表达改变对糖尿病大鼠心脏组织中蛋白表达谱的影响及可能的调节机制。方法:6周龄雄性RhoE基因敲除(KO)与野生型(WT)SD大鼠以腹腔一次性注射链脲佐菌素(STZ,70 mg/kg)复制急性1型糖尿病模型(T1DM),以注射等量柠檬酸钠盐水为对照,1周后连续3 d测定大鼠空腹血糖,以血糖浓度≥16.7 mmol/L为造模成功。2周后取出各组大鼠心脏,采用四维非标定量蛋白质组学技术(4D-LFQ),检测心脏组织中蛋白质的表达,筛选差异表达蛋白(DEP),生物信息学分析DEP相关功能和涉及的信号通路与蛋白-蛋白互作网络。结果:成功建立T1DM大鼠模型。以表达倍数(FC)>1.5倍且P<0.05为筛选标准,共鉴定到可定量的蛋白总数为2931个。非糖尿病状态下,KO组与WT组相比,上调表达蛋白26个,下调蛋白45个;糖尿病状态下,与WT组相比,KO组显著上调的蛋白数量为19个,显著下调的蛋白数量为28个。GO注释结果显示大多数差异表达蛋白位于细胞外基质区,生物学功能主要集中在免疫反应、能量代谢等过程。KEGG分析提示RhoE敲除后心脏组织中DEP参与的信号通路主要为核糖体、脂肪消化吸收等。蛋白互作网络分析发现KO组心脏组织中上调蛋白中Col1α1和Col1α2相互作用的蛋白较多,下调蛋白中参与核糖体通路的相关蛋白在网络互作较多。结论:RhoE敲除后明显改变了糖尿病心脏组织中蛋白质表达谱,影响多条与糖尿病心肌病关系密切的信号通路,研究结果为糖尿病心肌病的发病机制和治疗标靶筛选提供了参考。Objective:To analyze the effect of RhoE gene expression change on protein expression profiles in the cardiac tissue of diabetic rats,and analyse the possible underlying regulatory mechanism.Methods:Six-week-old male RhoE gene knockout(KO)and wild-type(WT)Sprague Dawley rats were injected intraperitoneally with streptozotocin(70 mg/kg)to establish the type 1 diabetes model(T1DM),with injection of the same amount of sodium citrate saline as the control group.A week later,the fasting blood glucose of the rats was measured daily,and blood glucose concentration≥16.7 mmol/L was regarded as a successful model.Two additional weeks later,the hearts of the rats in each group were removed and four-dimensional label-free quantitative proteomics technology(4D-LFQ)was used to analyze the changes of protein profiles in the heart tissue.The related functions,enrichment signals,and protein‐protein interaction network(PPI)of the differentially expressed proteins were analyzed.Results:T1DM rat models were successfully established.Taking a fold change>1.5 and P<0.05 as the threshold,a total of 2931 quantifiable proteins were identified.In the non-diabetic state,the KO group had 26 and 45 significantly up-and downregulated proteins,respectively,compared with the WT group;in the diabetic state,the KO group had 19 and 28 significantly up-and downregulated proteins,respectively,compared with the WT group.The GO annotation results showed that most of the differentially expressed proteins were located in the extracellular matrix,and their biological functions were mainly concentrated in the im‐mune response and energy metabolism.KEGG analysis showed that the signaling pathways associated with the differentially expressed proteins in cardiac tissue after RhoE knockout were mainly related to ribosomes and fat digestion and absorption.Protein interaction network analysis showed that in the cardiac tissue of the KO group,there were more Col1α1-and Col1α2-interacting proteins among the upregulated proteins,and among the down-regulated

关 键 词：糖尿病 心肌组织 RHOE 蛋白质组学 生物信息学 

分 类 号：R587.1[医药卫生—内分泌]