基于蛋白质组学探讨高糖致体外血脑屏障损伤的机制  

Mechanisms of high glucose-induced blood brain barrier injury in vitro using 4D-DIA technology

作　　者：张扬[1] 魏星 单昌友 赵永林 ZHANG Yang;WEI Xing;SHAN Changyou;ZHAO Yonglin(Department of Radiation Oncology,Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China;Department of Obstetrics and Gynecology,Second Affiliated Hospital of Xi'an Jiaotong University;Comprehensive Breast Care Center,Second Affiliated Hospital of Xi'an Jiaotong University)

机构地区：[1]西安交通大学第二附属医院肿瘤放疗科,西安710004 [2]西安交通大学第二附属医院妇产科 [3]西安交通大学第二附属医院乳腺疾病诊疗中心

出　　处：《山西医科大学学报》2025年第3期275-280,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(82001327)。

摘　　要：目的基于四维数据独立采集(4D-data-independent acquisition,4D-DIA)蛋白质组学技术,探究高糖致体外血脑屏障(blood brain barrier,BBB)损伤的机制。方法用小鼠脑微血管内皮细胞bEnd.3构建体外BBB模型,随机分为高糖组和对照组,高糖组接受含200 mmol/L葡萄糖的培养基处理24 h,对照组接受相同渗透压的含25 mmol/L葡萄糖的培养基处理24 h。检测跨内皮电阻(transepithelial resistance,TEER)及辣根过氧化物酶(horseradish peroxidase,HRP)评估体外BBB的通透性。利用4D-DIA技术检测并比较两组间差异蛋白的表达,对差异蛋白进行京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析。结果与对照组相比,高糖组HRP升高,TEER降低(均P<0.05)。两组共鉴定出603个差异蛋白,其中显著上调蛋白444个,显著下调蛋白159个。KEGG通路分类分析结果显示,差异蛋白主要富集于转运与分解代谢、信号转导、能量代谢、免疫系统等。KEGG信号通路分析显示,差异蛋白主要与氧化磷酸化、代谢等信号通路有关。结论高糖可能通过氧化磷酸化、代谢等信号通路影响内皮细胞的转运与分解代谢、信号转导、能量代谢、免疫系统等,进而增加体外BBB的通透性。Objective To investigate the mechanism of high glucose inducing blood brain barrier(BBB)damage in vitro using the fourdimensional data-independent acquisition(4D-DIA)proteomics technology.Methods The bEnd.3 mouse brain microvascular endothelial cells were used to induce an in vitro BBB model,and then randomly divided into high glucose group and control group.The cells in high glucose group were treated with medium containing 200 mmol/L glucose for 24 h,while the cells in control group were treated with medium containing 25 mmol/L glucose at the same osmotic pressure for 24 h.Transepithelial resistance(TEER)and horseradish peroxidase(HRP)were detected to evaluate BBB permeability in vitro.Differential protein expression between the two groups was detected and compared using 4D-DIA technology.The differential proteins were analyzed by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.Results Compared with control group,HRP was increased and TEER was decreased in high glucose group(both P<0.05).A total of 603 differential proteins were identified between the two groups,with 444 significantly upregulated proteins and 159 significantly downregulated proteins.KEGG pathway classification analysis indicated that the differential proteins were mainly concentrated in transport and catabolism,signal transduction,energy metabolism,immune system,etc.KEGG signaling pathway analysis showed that differential proteins were mainly related to oxidative phosphorylation,metabolism and other signaling pathways.Conclusion High glucose may increase the permeability of BBB in vitro by affecting the transport and catabolism,signal transduction,energy metabolism,and immune system of endothelial cells through oxidative phosphorylation and metabolism signaling pathways.

关 键 词：4D-DIA 蛋白质组学 高糖 血脑屏障 脑微管内皮细胞 通透性 

分 类 号：R363[医药卫生—病理学]